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Transesterification synthesis of high-yield biodiesel from black soldier fly larvae by using the combination of Lipase Eversa Transform 2.0 and Lipase SMG1

Abstract

The larvae of black soldier fly (BSFL) are rich in lipids, and in the current study, BSFL was used to produce biodiesel by enzymatic transesterification with methanol. Biodiesel obtained by free lipase-catalyzed transesterification is always with side-reactions. To address the limitation of free lipase, for the first time, Lipase SMG1 and Lipase Eversa Transform 2.0 were combined to use as catalysts at the same time. The effects of different reaction conditions on the yield of biodiesel were investigated, including the type of enzyme, reaction temperature, and molar ratio of BSFL to methanol. The molar ratio of BSFL to methanol 1:3, one-step addition of methanol at 25 °C were selected as optimal conditions. The fatty acid methyl ester content achieved to 98.45% after 8h reaction under the optimal conditions. Moreover, the main properties of the final biodiesel indicators met the EN14214 biodiesel standard.

Keywords:
enzyme; black soldier fly larvae; biodiesel

1 Introduction

Considering the environmental threats associated with the excessive use of fossil fuels (Yin et al., 2020Yin, Z. H., Zhu, L. D., Li, S. X., Hu, T. Y, Chu, R. Y, Mo, F., Hu, D., Liu, C. C, & Li, B. (2020). A comprehensive review on cultivation and harvesting of microalgae for biodiesel production: environmental pollution control and future directions. Bioresource Technology, 301, 122804. http://dx.doi.org/10.1016/j.biortech.2020.122804. PMid:31982297.
http://dx.doi.org/10.1016/j.biortech.202...
), it is crucial to explore alternative renewable energy sources. In recent years, biodiesel, produced from renewable sources such as vegetable oils, has attracted increasing attention as an alternative to traditional petroleum-based diesel. The main component of biodiesel is fatty acid methyl ester (FAME), which can be obtained from the transesterification of oils and alcohols (Bhatia et al., 2020Bhatia, S. K., Gurav, R., Choi, T. R., Kim, H. J., Yang, Y. H., Song, H. S., Park, J. Y., Park, Y. L., Han, Y. H., Choi, Y. K., Kim, S. H., Yoon, J. J., & Yang, Y. H. (2020). Conversion of waste cooking oil into biodiesel using heterogenous catalyst derived from cork biochar. Bioresource Technology, 302, 122872. http://dx.doi.org/10.1016/j.biortech.2020.122872. PMid:32014731.
http://dx.doi.org/10.1016/j.biortech.202...
; Costa et al., 2019Costa, W. A., Bezerra, F. W. F., Oliveira, M. S., Silva, M. P., Cunha, V. M. B., Andrade, E. H. A., & Carvalho, R. N. Jr. (2019). Appliance of a high pressure semi-batch reactor: supercritical transesterification of soybean oil using methanol. Food Science and Technology, 39(3), 754-773. http://dx.doi.org/10.1590/fst.05118.
http://dx.doi.org/10.1590/fst.05118...
). The commonly used feedstocks for biodiesel production include edible and non-edible oils which contain a large scale of triacylglycerols (TAG) (Knothe & Razon, 2017Knothe, G., & Razon, L. F. (2017). Biodiesel fuels. Progress in Energy and Combustion Science, 58, 36-59. http://dx.doi.org/10.1016/j.pecs.2016.08.001.
http://dx.doi.org/10.1016/j.pecs.2016.08...
). However, using edible oils in biodiesel production is costly and may compete with food supply, thus rendering biodiesel less competitive in the fuel market (Nguyen et al., 2017Nguyen, H. C., Liang, S. H, Doan, T. T, Su, C. H, & Yang, P. C. (2017). Lipase-catalyzed synthesis of biodiesel from black soldier fly (Hermetica illucens): optimization by using response surface methodology. Energy Conversion and Management, 145, 335-342. http://dx.doi.org/10.1016/j.enconman.2017.05.010.
http://dx.doi.org/10.1016/j.enconman.201...
). Using non-edible oils and waste cooking oils can largely reduce the cost in biodiesel production, but these oils usually require additional processing steps. Therefore, it is necessary to explore a new and cost-effective source of oils for biodiesel production to reduce the cost and simplify the process (Surendra et al., 2016Surendra, K. C., Olivier, R., Tomberlin, J. K., Jha, R., & Khanal, S. K. (2016). Bioconversion of organic wastes into biodiesel and animal feed via insect farming. Renewable Energy, 98, 197-202. http://dx.doi.org/10.1016/j.renene.2016.03.022.
http://dx.doi.org/10.1016/j.renene.2016....
).

The black soldier fly is a bi-winged insect based of the soldier fly family. Unlike other insects, the black soldier fly is not a pest and can survive in a variety of complex environments such as kitchen waste and poultry manure with a short growth cycle (Somroo et al., 2019Somroo, A. A., Rehman, K. U., Zheng, L., Cai, M., Xiao, X., Hu, S., Mathys, A., Gold, M., Yu, Z., & Zhang, J. (2019). Influence of lactobacillus buchneri on soybean curd residue co-conversion by black soldier fly larvae (hermetia illucens) for food and feedstock production. Waste Management, 86, 114-122. http://dx.doi.org/10.1016/j.wasman.2019.01.022. PMid:30902235.
http://dx.doi.org/10.1016/j.wasman.2019....
). More importantly, it can inactivate pathogens and convert a large amount of low-value organic waste to high-value biomass, which makes them quite popular in many areas like waste-reducing (Singh & Kumari, 2019Singh, A., & Kumari, K. C. (2019). An inclusive approach for organic waste treatment and valorisation using Black Soldier Fly larvae: a review. Journal of Environmental Management, 251, 109569. http://dx.doi.org/10.1016/j.jenvman.2019.109569. PMid:31550603.
http://dx.doi.org/10.1016/j.jenvman.2019...
). The larvae of black soldier fly (BSFL) are rich of oils and proteins, containing 35-40% and 40-44% of lipids and crude proteins, respectively. The extracted lipids from BSFL are mainly composed of saturated fatty acids (SFFA) and have been used as in the production of biodiesel (Wang et al., 2017aWang, C. W., Qian, L., Wang, W. G., Wang, T. L., Deng, Z. K., Yang, F., Xiong, J., & Feng, W. L. (2017a). Exploring the potential of lipids from black soldier fly: new paradigm for biodiesel production (I). Renewable Energy, 111, 749-756. http://dx.doi.org/10.1016/j.renene.2017.04.063.
http://dx.doi.org/10.1016/j.renene.2017....
; Cutrignelli et al., 2018Cutrignelli, M. I., Messina, M., Tulli, F., Randazzo, B., Olivotto, I., Gasco, L., Loponte, R., & Bovera, F. (2018). Evaluation of an insect meal of the black soldier fly (hermetia illucens) as soybean substitute: intestinal morphometry, enzymatic and microbial activity in laying hens. Research in Veterinary Science, 117, 209-215. http://dx.doi.org/10.1016/j.rvsc.2017.12.020. PMid:29304440.
http://dx.doi.org/10.1016/j.rvsc.2017.12...
; Ewald et al., 2020Ewald, N., Vidakovic, A., Langeland, M., Kiessling, A., Sampels, S., & Lalander, C. (2020). Fatty acid composition of black soldier fly larvae (hermetia illucens) – possibilities and limitations for modification through diet. Waste Management, 102, 40-47. http://dx.doi.org/10.1016/j.wasman.2019.10.014. PMid:31655329.
http://dx.doi.org/10.1016/j.wasman.2019....
). Therefore, lipids extracted from BSFL represent a potential source for biodiesel production.

Compared with chemical catalysts, enzymes require a mild reaction environment and be able to display specific selectivity during transesterification and thus can produce biodiesel with limited by-products in an environmentally friendly way (Moazeni et al., 2019Moazeni, F., Chen, Y. C., & Zhang, G. (2019). Enzymatic transesterification for biodiesel production from used cooking oil, a review. Journal of Cleaner Production, 216(2), 117-128. http://dx.doi.org/10.1016/j.jclepro.2019.01.181.
http://dx.doi.org/10.1016/j.jclepro.2019...
). The type of catalysts plays a crucial role in the yield of biodiesel (Martínez-Corona et al., 2020Martínez-Corona, R., Banderas-Martínez, F. J., Pérez-Castillo, J. N., Cortés-Penagos, C., & González-Hernández, J. C. (2020). Avocado oil as an inducer of the extracellular lipase activity of kluyveromyces marxianus l-2029. Food Science and Technology, 40(Suppl. 1), 121-129. http://dx.doi.org/10.1590/fst.06519.
http://dx.doi.org/10.1590/fst.06519...
). There are two main types of enzymes used for transesterification: immobilized lipase and free lipase. The immobilized lipase has many advantages including reusability, stability, and high yield etc. (Wang et al., 2017b; Cardoso et al., 2021Cardoso, F. S. N., Carvalho, L. M. J. D., Koblitz, M. G. B., & Ortiz, G. M. D. (2021). Use of encapsulated commercial enzyme in the hydrolysis optimization of cagaita pulp (eugenia dysenterica dc). Food Science and Technology. Ahead of print.). For instance, Talukder et al. (2011)Talukder, M., Das, P., Tan, S. F., & Jin, C. W. (2011). Enhanced enzymatic transesterification of palm oil to biodiesel. Biochemical Engineering Journal, 55(2), 119-122. used Novozym 435 as catalyst for transesterification of palm oil and the yield of biodiesel reached up to 95-96% after 8 h of reaction. However, the commercial immobilized lipase usually costs a lot, while free lipase costs much lower than it. For instance, the price of immobilized lipase like Novozyme 435, is 2286.00 $/kg while the cost of Lipase Eversa Transform 2.0 is 150.00 $/kg only (Sun et al., 2021Sun, S. D., Guo, J. J., & Chen, X. W. (2021). Biodiesel preparation from Semen Abutili (Abutilon theophrasti Medic.) seed oil using low-cost liquid lipase Eversa® transform 2.0 as a catalyst. Industrial Crops & Products, 169, 113643.). But, the disadvantages of free lipase are also obvious including higher acid value, longer reaction time and higher content of unpleasant by-product. Thus, molecular distillation will be used to purify biodiesel, which means the cost will be more expensive and the process is complicated (Lv et al., 2021Lv, L., Dai, L., Du, W., & Liu, D. (2021). Progress in enzymatic biodiesel production and commercialization. Processes, 9(2), 355. http://dx.doi.org/10.3390/pr9020355.
http://dx.doi.org/10.3390/pr9020355...
). Therefore, the catalysts and the process of biodiesel prepared still need to be improved with the reduction of cost and the increase of yield. Generally, monoacylglycerol (MAG) and diacylglycerol (DAG) lipases are used to remove partial glycerides from glyceride mixtures (Li et al., 2015Li, D. M., Qin, X. L., Wang, J. R., Yang, B., Wang, W. F., Huang, W. L., & Wang, Y. H. (2015). Hydrolysis of soybean oil to produce diacylglycerol by a lipase from Rhizopus -oryzae. Journal of Molecular Catalysis. B, Enzymatic, 115, 43-50. http://dx.doi.org/10.1016/j.molcatb.2015.01.009.
http://dx.doi.org/10.1016/j.molcatb.2015...
). Many efforts had been done in the catalytic ability of Lipase SMG1, and it’s the first time Lipase SMG1 was used in the production of biodiesel to obtain lower content of fatty acid (FFA), DAG, and MAG in the final system.

The aim of this study, therefore, is to develop a cost-effective and simple approach to produce a high yield of biodiesel using BSFL lipids and enzymes. The effect of enzyme types, molar ratio of BSFL lipids to methanol, and reaction temperature on biodiesel production were investigated. The properties of the final biodiesel products were further compared with literature and the EN14214 biodiesel standard.

2 Materials and methods

2.1 Materials

The BSFL lipids, a by-product from black soldier fly defatted meal factory, were kindly supplied by Guangzhou Fishtech Technology Co. Ltd. (Guangdong, PR China). Briefly, ground larval biomass was placed into a filter bag and soaked in petroleum ether for 48 h twice at room temperature. Lipase Eversa Transform 2.0, was purchased from Novozymes (Tianjin, China). Lipase SMG1 (free lipase) was produced according to the method of Xu et al. (2012)Xu, T. T, Liu, L., Hou, S. L., Xu, J. X., Yang, B., Wang, Y. H., & Liu, J. S. (2012). Crystal structure of a mono- and diacylglycerol lipase from Malassezia globosa reveals a novel lid conformation and insights into the substrate specificity. Journal of Structural Biology, 178(3), 363-369. http://dx.doi.org/10.1016/j.jsb.2012.03.006. PMid:22484238.
http://dx.doi.org/10.1016/j.jsb.2012.03....
. Lipase G50 (free lipase) was purchased from Amano Enzyme (Japan). Methanol (Analytically pure grade) was used as an acyl acceptor, other chemicals (n-hexane, iso-propanol et al.) were all HPLC grade. All chemicals were obtained from Sigma Aldrich (Shanghai, China).

2.2 Preparation of biodiesel by enzyme-catalyzed transesterification

Lipase Eversa Transform 2.0 alone or in combination with lipases SMG1 (or Lipase G50) were used as enzyme catalysts for transesterification of BSFL lipids and their efficiencies in the production of biodiesel were compared. The effects of molar ratio of BSFL lipids to methanol (1:2, 1:3, 1:4, 1:5) and reaction temperature (20, 25, 30, 35 °C) on the yield of biodiesel were investigated, respectively. Samples were collected periodically (every 2 h) till 10 h of reaction and were analyzed by high-performance liquid chromatography (HPLC).

2.3 Analysis of the composition of the reaction mixtures by HPLC

The analysis of the composition of the reaction mixtures was carried out using a normal-phase HPLC equipped with a refractive index detector and a Phenomenex Luna column (250 mM × 4.6 mM i.d., 5 μm particle size) according to the method previously described by Li et al. (2015)Li, D. M., Qin, X. L., Wang, J. R., Yang, B., Wang, W. F., Huang, W. L., & Wang, Y. H. (2015). Hydrolysis of soybean oil to produce diacylglycerol by a lipase from Rhizopus -oryzae. Journal of Molecular Catalysis. B, Enzymatic, 115, 43-50. http://dx.doi.org/10.1016/j.molcatb.2015.01.009.
http://dx.doi.org/10.1016/j.molcatb.2015...
with a minor modification. The mobile phase consisted of n-hexane, iso-propanol and formic acid (21:1:0.003, by vol) with a flow rate of 1 mL/min. Peaks in HPLC were evaluated by comparison of their retention times with those known standards.

The yield of FAME was calculated follows the Equation 1:

F A M E c o n t e n t % = FAME % TAG % + DAG % + MAG % + FFA % + FAME % × 100 % (1)

2.4 Analysis of fatty acid composition by GC

The final biodiesel was initially methylated to FAME according to the method described by Wang et al. (2010)Wang, Y. H., Mai, Q. Y., Qin, X. L., Yang, B., Wang, Z. L., & Chen, H. T. (2010). Establishment of an evaluation model for human milk fat substitutes. Journal of Agricultural and Food Chemistry, 58(1), 642-649. http://dx.doi.org/10.1021/jf903048p. PMid:20000702.
http://dx.doi.org/10.1021/jf903048p...
. Then, FFA composition of the final FAME product was determined using GC (Agilent 7890A) equipped with a capillary column CP-Sil 88 (60 mM × 0.25 mM,0.2 μm). The analysis was carried out according to the method described by Qin et al. (2011)Qin, X. L., Wang, X. M., Wang, Y. H., Huang, H. H., & Yang, B. (2011). Preparation and characterization of 1,3-dioleoyl-2-palmitoylglycerol. Journal of Agricultural and Food Chemistry, 59(10), 5714-5719. http://dx.doi.org/10.1021/jf200398f. PMid:21510711.
http://dx.doi.org/10.1021/jf200398f...
.

2.5 Analysis density, viscosity, flash point and acid value of biodiesel

The density, viscosity, flash point, and acid value of the final biodiesel product were all tested according to the methods of the American Society for Testing and Materials (ASTM).

2.6 Statistical analysis

All experiments were carried in triplicate. The results were presented as the means ± standard deviations (SD). The statistical significance of differences was measured by a one-way analysis of variance (ANOVA).

3 Results and discussion

3.1 The effect of enzyme type on the yield of biodiesel

Lipase Eversa Transform 2.0 alone or in combination with Lipase SMG1 (or Lipase G50) were used as catalysts for transesterification. As illustrated in Figure 1, the yield of FAME reached up to 66.74% when Lipase Eversa Transform 2.0 alone was used as a catalyst, and even didn’t reach a reaction equilibrium after 8h. The low-quality of BSFL may make this phenomenon occurred, and the raw materials may have phospholipids and unpurified contents. Li et al. (2014)Li, Y., Du, W., & Liu, D. (2014). Exploration on the effect of phospholipids on free lipase-mediated biodiesel production. Journal of Molecular Catalysis. B, Enzymatic, 102, 88-93. http://dx.doi.org/10.1016/j.molcatb.2014.01.018.
http://dx.doi.org/10.1016/j.molcatb.2014...
had reported that when the content of phospholipids exceeds 5%, the reaction rate will be decreased. However, the FAME yield increased dramatically when using the combination of Lipase Eversa Transform 2.0 and Lipase SMG1/LipaseG50 as the catalysts.

Figure 1
Effect of enzyme types on FAME content. (The transesterification was carried out at 25 °C with a molar ratio 1:3 between BSFL lipids and methanol, the lipase dosage was 50 U/g Lipase Eversa Transform 2.0 alone or 50 U/g Lipase Eversa Transform 2.0 combined with 50 U/g Lipase SMG1/ Lipase G50.)

A yield of 98.45% was obtained in the presence of both Lipase Eversa Transform 2.0 and Lipase SMG1.Figure 2 illustrated that after the transesterification, the content of TAG (1.02%), DAG (0.26%), MAG (0.10%), and FFA (0.17%) occupied less than that of the raw materials, and there was a high yield of FAME (98.45%). Lipase Eversa Transform 2.0 is a TAG lipase that catalyzes TAG hydrolysis only (Remonatto et al., 2018Remonatto, D., Oliveira, J. V., Guisan, J. M., Oliveira, D., Ninow, J., & Fernandez-Lorente, G. (2018). Production of fame and faee via alcoholysis of sunflower oil by eversa lipases immobilized on hydrophobic supports. Applied Biochemistry and Biotechnology, 185(3), 705-716. http://dx.doi.org/10.1007/s12010-017-2683-1. PMid:29297136.
http://dx.doi.org/10.1007/s12010-017-268...
) and Lipase SMG1 and Lipase G50 are partial glyceride lipases that have catalytic activity towards DAG and monoacylglycerol (MAG) according to Xu et al. (2012)Xu, T. T, Liu, L., Hou, S. L., Xu, J. X., Yang, B., Wang, Y. H., & Liu, J. S. (2012). Crystal structure of a mono- and diacylglycerol lipase from Malassezia globosa reveals a novel lid conformation and insights into the substrate specificity. Journal of Structural Biology, 178(3), 363-369. http://dx.doi.org/10.1016/j.jsb.2012.03.006. PMid:22484238.
http://dx.doi.org/10.1016/j.jsb.2012.03....
and Zheng et al. (2014)Zheng, P. Y., Xu, Y., Wang, W. F., Qin, X. L., Ning, Z. X., Wang, Y. H., & Yang, B. (2014). Production of diacylglycerol-mixture of regioisomers with high purity by two-step enzymatic reactions combined with molecular distillation. Journal of the American Oil Chemists’ Society, 91(2), 251-259. http://dx.doi.org/10.1007/s11746-013-2365-2.
http://dx.doi.org/10.1007/s11746-013-236...
. Particularly, the Lipase SMG1 showed well catalyzed ability between FFA and methanol which has positive effect in increasing the extent of reaction and decreasing the acid value obviously (Li et al., 2016Li, D. M., Wang, W. F., Durrani, R., Li, X. X., Yang, B., & Wang, Y. H. (2016). Simplified enzymatic upgrading of high-acid rice bran oil using ethanol as a novel acyl acceptor. Journal of Agricultural and Food Chemistry, 64(35), 6730-6737. http://dx.doi.org/10.1021/acs.jafc.6b02518. PMid:27571030.
http://dx.doi.org/10.1021/acs.jafc.6b025...
). In this study, after transesterification, the content of FFA dropped significantly from 11.35% to 0.17%, and the acid value was about 0.10 mg KOH/g. Therefore, the increase of FAME yield likely caused by the effective transesterification upon all types of glycerides, FFA, and even phospholipids which was consistent with the catalytic characteristics of Lipase Eversa Transform 2.0 and Lipase SMG1 mentioned before. Thus, the mixture of Lipase Eversa Transform 2.0 and Lipase SMG1 was used as catalysts in further study.

Figure 2
The composition of BSFL and biodiesel detected by HPLC.

3.2 The effect of reaction temperature on the yield of biodiesel

Figure 3 showed the effect of reaction temperature on the yield of FAME. The yield of biodiesel increased from 73.90% to 98.45% with the reaction temperature increased from 20 to 25 °C. When the temperature was further increased to 30 °C, although the initial yield in first 2 h was higher comparable to that at 25 °C but the final yield dropped to 86.43% after 10 h. The FAME production dropped markedly when the reaction temperature reached to 35 °C. This is likely because of a dual effect of temperature on enzyme catalytic activity, where a higher reaction rate and yield are obtained at a higher temperature, while increasing the temperature beyond certain point (30 °C) leads to enzyme inactivation. Therefore, the reaction temperature was set at 25 °C in the following study.

Figure 3
Effect of reaction temperature on the yield of FAME. (Lipase Eversa Transform 2.0 together with Lipase SMG1 were used as catalysts. Transesterification was performed with a molar ratio of BSFL lipids to methanol 1:3 and one-step addition methanol at varying temperature (20, 25, 30, and 35 °C).)

3.3 The effect of the substrate molar ratio on the yield of biodiesel

The effect of substrate molar ratio on FAME production was determined by setting the molar ratio of BSFL lipids to methanol at 1:2, 1:3, 1:4, and 1:5, respectively, in the transesterification. The results were showed in Figure 4. When the molar ratio of BSFL lipids to methanol increased from 1:2 to 1:3, the yield of FAME achieved up to 98.45%. A further increase of the molar ratio of BSFL lipids to methanol beyond 1:3 led to the yield of biodiesel decreased significantly. In the transesterification reaction, the ideal molar ratio of oil to methanol is 1:3. Increasing the molar ratio will make the reaction move forward and accelerate the reaction process. However, the short-chain alcohol has a toxic effect on the enzyme activity (Xu et al., 2012Xu, T. T, Liu, L., Hou, S. L., Xu, J. X., Yang, B., Wang, Y. H., & Liu, J. S. (2012). Crystal structure of a mono- and diacylglycerol lipase from Malassezia globosa reveals a novel lid conformation and insights into the substrate specificity. Journal of Structural Biology, 178(3), 363-369. http://dx.doi.org/10.1016/j.jsb.2012.03.006. PMid:22484238.
http://dx.doi.org/10.1016/j.jsb.2012.03....
), resulting in a notable reduction on the final product content (Su’i et al., 2021Su’i, M., Sumaryati, E., Dwianggraeni, F., Utomo, Y., Anggraini, N., & Rofiqoh, N. L. (2021). Monoglyceride biosynthesis from coconut milk with lypase enzyme of sesame seed sprouts as biocatalyst. Food Science and Technology, 41(Suppl. 1), 328-333.). Therefore, the optimal condition for FAME production was established to be a molar ratio of BSFL lipids to methanol at 1:3, reaction time at 25 °C and a mixture of Lipase Eversa Transform 2.0 and Lipase SMG1 as catalysis. Under this condition, a maximum FAME yield of 98.45% was obtained after 8 h.

Figure 4
Effect of the molar ratio of BSFL to methanol on the yield of FAME. (Lipase Eversa Transform 2.0 together with Lipase SMG1 were used as catalysts. Transesterification was performed at 25°C, with a molar ratio of BSFL to methanol at 1:2, 1:3, 1:4, or 1:5.)

3.4 Properties of biodiesel

The cetane index, viscosity, ester content, acid value, and flash point are important properties of biodiesel. And the cetane index, viscosity, and flash point are related with carbon number, molar weight, and unsaturated carbon bonds number, etc. (Giakoumis & Sarakatsanis, 2018Giakoumis, E. G., & Sarakatsanis, C. K. (2018). Estimation of biodiesel cetane number, density, kinematic viscosity and heating values from its fatty acid weight composition. Fuel, 222, 574-585. http://dx.doi.org/10.1016/j.fuel.2018.02.187.
http://dx.doi.org/10.1016/j.fuel.2018.02...
). Table 1 and Figure 5 compared the composition of FFA among the final biodiesel product obtained in this study, the biodiesel produced from BSFL using Novozym 435 as catalyst (Nguyen et al., 2018Nguyen, H. C., Liang, S. H., Li, S., Su, C., Chien, C., Chen, Y., & Huong, D. T. M. (2018). Direct transesterification of black soldier fly larvae (Hermetia illucens) for biodiesel production. Journal of the Taiwan Institute of Chemical Engineers, 85, 165-169. http://dx.doi.org/10.1016/j.jtice.2018.01.035.
http://dx.doi.org/10.1016/j.jtice.2018.0...
) and palm oil (Sajjadi et al., 2016Sajjadi, B., Raman, A., & Arandiyan, H. (2016). A comprehensive review on properties of edible and non-edible vegetable oil-based biodiesel: composition, specifications and prediction models. Renewable & Sustainable Energy Reviews, 63, 62-92. http://dx.doi.org/10.1016/j.rser.2016.05.035.
http://dx.doi.org/10.1016/j.rser.2016.05...
) reported in the literatures before. And the main properties of the final biodiesel produced in this study and reported in the literatures were compared according to the standard EN14214 in Table2.

Table 1
The fatty acid composition of the biodiesel produced from BSFL, palm oil and this study.
Figure 5
The comparison of UFFA and SFFA content among different type of biodiesel.
Table 2
Main properties of different biodiesel types compared.

Although the raw materials of this study and the research reported by Nguyen et al. (2018)Nguyen, H. C., Liang, S. H., Li, S., Su, C., Chien, C., Chen, Y., & Huong, D. T. M. (2018). Direct transesterification of black soldier fly larvae (Hermetia illucens) for biodiesel production. Journal of the Taiwan Institute of Chemical Engineers, 85, 165-169. http://dx.doi.org/10.1016/j.jtice.2018.01.035.
http://dx.doi.org/10.1016/j.jtice.2018.0...
were both BSFL, the fatty acid composition of them had significant difference. Many studies have showed that the nutrient content of a BSFL product including lipids composition is highly related with the growing substrate of BSFL (Tschirner & Simon, 2015Tschirner, M., & Simon, A. (2015). Influence of different growing substrates and processing on the nutrient composition of black soldier fly larvae destined for animal feed. Journal of Insects as Food and Feed, 1(4), 249- 259. http://dx.doi.org/10.3920/JIFF2014.0008.
http://dx.doi.org/10.3920/JIFF2014.0008...
). And the BSFL biodiesel reported before had a lower unsaturated fatty acids (UFFA) content (33.50%) than that of this study (50.26%), which resulted a lower cetane index, higher viscosity, and flash point. This phenomenon was consistence with the relation between properties and FFA composition. The kinematic viscosity decreases while the cetane index increases as the total content of UFFA increases. (Giakoumis & Sarakatsanis, 2018Giakoumis, E. G., & Sarakatsanis, C. K. (2018). Estimation of biodiesel cetane number, density, kinematic viscosity and heating values from its fatty acid weight composition. Fuel, 222, 574-585. http://dx.doi.org/10.1016/j.fuel.2018.02.187.
http://dx.doi.org/10.1016/j.fuel.2018.02...
; Kaisan et al, 2017Kaisan, M. U., Anafi, F. O., Nuszkowski, J., Kulla, D. M., & Umaru, S. (2017). Calorific value, flash point and cetane number of biodiesel from cotton, jatropha and neem binary and multi-blends with diesel. Biofuels, 11(3), 321-327.). The flash point decreases with increasing of residual alcohol and other solvents with a low-boiling point (Černoch et al, 2010Černoch, M., Hájek, M., & Skopal, F. (2010). Relationships among flash point, carbon residue, viscosity and some impurities in biodiesel after ethanolysis of rapeseed oil. Bioresource Technology, 101(19), 7397-7401. http://dx.doi.org/10.1016/j.biortech.2010.05.003. PMid:20537532.
http://dx.doi.org/10.1016/j.biortech.201...
). Besides, higher flash point value means better for biodiesel handling, storage, and transportation (Alviso et al., 2020Alviso, D., Saab, E. E., Clevenot, P., & Romano, S. D. (2020). Flash point, kinematic viscosity and refractive index: variations and correlations of biodiesel–diesel blends. Journal of the Brazilian Society of Mechanical Sciences and Engineering, 42(6), 347. http://dx.doi.org/10.1007/s40430-020-02428-w.
http://dx.doi.org/10.1007/s40430-020-024...
).

The biodiesel obtained in the current study has 49.74% SFFA and 50.26% UFFA totally, which had no significant difference from the biodiesel prepared by palm oil. Despite this, the cetane number, viscosity, and flash point between them still had difference, this may be related to the SFFA composition of them was different like BSFL biodiesel of this study had more lauric and myristic. What’s more, the quality of raw material and processing conditions during the preparation of biodiesel also effects the properties of biodiesel. (Ramirez-Verduzco et al., 2012Ramirez-Verduzco, L. F., Rodriguez-Rodriguez, J. E., & Jaramillo-Jacob, A. (2012). Predicting cetane number, kinematic viscosity, density and higher heating value of biodiesel from its fatty acid methyl ester composition. Fuel, 91(1), 102-111. http://dx.doi.org/10.1016/j.fuel.2011.06.070.
http://dx.doi.org/10.1016/j.fuel.2011.06...
).

The final biodiesel obtained in this study meets the requirements of the EN14214 standard, and has similar properties to biodiesel produced from palm oil. Moreover, the acid value was 0.10 mg KOH/g which was lower than the limitation of EN 14214 (0.50 mg KOH/g), thus shows great potential in replacing diesel usage in industry.

4 Conclusions

Here, we reported a cost-effective and simple method to produce biodiesel using BSFL lipids and enzyme catalysis. The results showed that using Lipase Eversa Transform 2.0 with Lipase SMG1 as catalyst was able to increase the yield of biodiesel and drop the acid value significantly. After optimization of the reaction condition, the highest FAME yield of 98.45% was achieved at 25 °C with a molar ratio of BSFL to methanol at 1:3. And the properties of the obtained biodiesel met the EN14214 standard. This study showed the combination of Lipase Eversa Transform 2.0 with Lipase SMG1 has great value in the production of biodiesel.

  • Practical Application: Lipase SMG1 and Lipase Eversa Transform 2.0 were combined to prepare biodiesel from black soldier fly larvae, which reduced the restrictions of free lipase.
  • Funding

    We gratefully acknowledge the financial support from the National Key R&D Program of China (2019YFD1002403), National Science Fund for Key Program of National Natural Science Foundation of China (31930084), Distinguished Young Scholars of China (31725022), China Agriculture Research System (CARS-18-ZJ0503), Science and Technology Planning Project of Guangdong Province (2019A050503002), Innovation and Entrepreneurship Team of Nanhai Talent Plan of Lanhai District, Foshan (201811070001).

References

  • Alviso, D., Saab, E. E., Clevenot, P., & Romano, S. D. (2020). Flash point, kinematic viscosity and refractive index: variations and correlations of biodiesel–diesel blends. Journal of the Brazilian Society of Mechanical Sciences and Engineering, 42(6), 347. http://dx.doi.org/10.1007/s40430-020-02428-w
    » http://dx.doi.org/10.1007/s40430-020-02428-w
  • Bhatia, S. K., Gurav, R., Choi, T. R., Kim, H. J., Yang, Y. H., Song, H. S., Park, J. Y., Park, Y. L., Han, Y. H., Choi, Y. K., Kim, S. H., Yoon, J. J., & Yang, Y. H. (2020). Conversion of waste cooking oil into biodiesel using heterogenous catalyst derived from cork biochar. Bioresource Technology, 302, 122872. http://dx.doi.org/10.1016/j.biortech.2020.122872 PMid:32014731.
    » http://dx.doi.org/10.1016/j.biortech.2020.122872
  • Cardoso, F. S. N., Carvalho, L. M. J. D., Koblitz, M. G. B., & Ortiz, G. M. D. (2021). Use of encapsulated commercial enzyme in the hydrolysis optimization of cagaita pulp (eugenia dysenterica dc). Food Science and Technology Ahead of print.
  • Černoch, M., Hájek, M., & Skopal, F. (2010). Relationships among flash point, carbon residue, viscosity and some impurities in biodiesel after ethanolysis of rapeseed oil. Bioresource Technology, 101(19), 7397-7401. http://dx.doi.org/10.1016/j.biortech.2010.05.003 PMid:20537532.
    » http://dx.doi.org/10.1016/j.biortech.2010.05.003
  • Costa, W. A., Bezerra, F. W. F., Oliveira, M. S., Silva, M. P., Cunha, V. M. B., Andrade, E. H. A., & Carvalho, R. N. Jr. (2019). Appliance of a high pressure semi-batch reactor: supercritical transesterification of soybean oil using methanol. Food Science and Technology, 39(3), 754-773. http://dx.doi.org/10.1590/fst.05118
    » http://dx.doi.org/10.1590/fst.05118
  • Cutrignelli, M. I., Messina, M., Tulli, F., Randazzo, B., Olivotto, I., Gasco, L., Loponte, R., & Bovera, F. (2018). Evaluation of an insect meal of the black soldier fly (hermetia illucens) as soybean substitute: intestinal morphometry, enzymatic and microbial activity in laying hens. Research in Veterinary Science, 117, 209-215. http://dx.doi.org/10.1016/j.rvsc.2017.12.020 PMid:29304440.
    » http://dx.doi.org/10.1016/j.rvsc.2017.12.020
  • Ewald, N., Vidakovic, A., Langeland, M., Kiessling, A., Sampels, S., & Lalander, C. (2020). Fatty acid composition of black soldier fly larvae (hermetia illucens) – possibilities and limitations for modification through diet. Waste Management, 102, 40-47. http://dx.doi.org/10.1016/j.wasman.2019.10.014 PMid:31655329.
    » http://dx.doi.org/10.1016/j.wasman.2019.10.014
  • Giakoumis, E. G., & Sarakatsanis, C. K. (2018). Estimation of biodiesel cetane number, density, kinematic viscosity and heating values from its fatty acid weight composition. Fuel, 222, 574-585. http://dx.doi.org/10.1016/j.fuel.2018.02.187
    » http://dx.doi.org/10.1016/j.fuel.2018.02.187
  • Kaisan, M. U., Anafi, F. O., Nuszkowski, J., Kulla, D. M., & Umaru, S. (2017). Calorific value, flash point and cetane number of biodiesel from cotton, jatropha and neem binary and multi-blends with diesel. Biofuels, 11(3), 321-327.
  • Knothe, G., & Razon, L. F. (2017). Biodiesel fuels. Progress in Energy and Combustion Science, 58, 36-59. http://dx.doi.org/10.1016/j.pecs.2016.08.001
    » http://dx.doi.org/10.1016/j.pecs.2016.08.001
  • Li, D. M., Qin, X. L., Wang, J. R., Yang, B., Wang, W. F., Huang, W. L., & Wang, Y. H. (2015). Hydrolysis of soybean oil to produce diacylglycerol by a lipase from Rhizopus -oryzae. Journal of Molecular Catalysis. B, Enzymatic, 115, 43-50. http://dx.doi.org/10.1016/j.molcatb.2015.01.009
    » http://dx.doi.org/10.1016/j.molcatb.2015.01.009
  • Li, D. M., Wang, W. F., Durrani, R., Li, X. X., Yang, B., & Wang, Y. H. (2016). Simplified enzymatic upgrading of high-acid rice bran oil using ethanol as a novel acyl acceptor. Journal of Agricultural and Food Chemistry, 64(35), 6730-6737. http://dx.doi.org/10.1021/acs.jafc.6b02518 PMid:27571030.
    » http://dx.doi.org/10.1021/acs.jafc.6b02518
  • Li, Y., Du, W., & Liu, D. (2014). Exploration on the effect of phospholipids on free lipase-mediated biodiesel production. Journal of Molecular Catalysis. B, Enzymatic, 102, 88-93. http://dx.doi.org/10.1016/j.molcatb.2014.01.018
    » http://dx.doi.org/10.1016/j.molcatb.2014.01.018
  • Lv, L., Dai, L., Du, W., & Liu, D. (2021). Progress in enzymatic biodiesel production and commercialization. Processes, 9(2), 355. http://dx.doi.org/10.3390/pr9020355
    » http://dx.doi.org/10.3390/pr9020355
  • Martínez-Corona, R., Banderas-Martínez, F. J., Pérez-Castillo, J. N., Cortés-Penagos, C., & González-Hernández, J. C. (2020). Avocado oil as an inducer of the extracellular lipase activity of kluyveromyces marxianus l-2029. Food Science and Technology, 40(Suppl. 1), 121-129. http://dx.doi.org/10.1590/fst.06519
    » http://dx.doi.org/10.1590/fst.06519
  • Moazeni, F., Chen, Y. C., & Zhang, G. (2019). Enzymatic transesterification for biodiesel production from used cooking oil, a review. Journal of Cleaner Production, 216(2), 117-128. http://dx.doi.org/10.1016/j.jclepro.2019.01.181
    » http://dx.doi.org/10.1016/j.jclepro.2019.01.181
  • Nguyen, H. C., Liang, S. H., Li, S., Su, C., Chien, C., Chen, Y., & Huong, D. T. M. (2018). Direct transesterification of black soldier fly larvae (Hermetia illucens) for biodiesel production. Journal of the Taiwan Institute of Chemical Engineers, 85, 165-169. http://dx.doi.org/10.1016/j.jtice.2018.01.035
    » http://dx.doi.org/10.1016/j.jtice.2018.01.035
  • Nguyen, H. C., Liang, S. H, Doan, T. T, Su, C. H, & Yang, P. C. (2017). Lipase-catalyzed synthesis of biodiesel from black soldier fly (Hermetica illucens): optimization by using response surface methodology. Energy Conversion and Management, 145, 335-342. http://dx.doi.org/10.1016/j.enconman.2017.05.010
    » http://dx.doi.org/10.1016/j.enconman.2017.05.010
  • Qin, X. L., Wang, X. M., Wang, Y. H., Huang, H. H., & Yang, B. (2011). Preparation and characterization of 1,3-dioleoyl-2-palmitoylglycerol. Journal of Agricultural and Food Chemistry, 59(10), 5714-5719. http://dx.doi.org/10.1021/jf200398f PMid:21510711.
    » http://dx.doi.org/10.1021/jf200398f
  • Ramirez-Verduzco, L. F., Rodriguez-Rodriguez, J. E., & Jaramillo-Jacob, A. (2012). Predicting cetane number, kinematic viscosity, density and higher heating value of biodiesel from its fatty acid methyl ester composition. Fuel, 91(1), 102-111. http://dx.doi.org/10.1016/j.fuel.2011.06.070
    » http://dx.doi.org/10.1016/j.fuel.2011.06.070
  • Remonatto, D., Oliveira, J. V., Guisan, J. M., Oliveira, D., Ninow, J., & Fernandez-Lorente, G. (2018). Production of fame and faee via alcoholysis of sunflower oil by eversa lipases immobilized on hydrophobic supports. Applied Biochemistry and Biotechnology, 185(3), 705-716. http://dx.doi.org/10.1007/s12010-017-2683-1 PMid:29297136.
    » http://dx.doi.org/10.1007/s12010-017-2683-1
  • Sajjadi, B., Raman, A., & Arandiyan, H. (2016). A comprehensive review on properties of edible and non-edible vegetable oil-based biodiesel: composition, specifications and prediction models. Renewable & Sustainable Energy Reviews, 63, 62-92. http://dx.doi.org/10.1016/j.rser.2016.05.035
    » http://dx.doi.org/10.1016/j.rser.2016.05.035
  • Singh, A., & Kumari, K. C. (2019). An inclusive approach for organic waste treatment and valorisation using Black Soldier Fly larvae: a review. Journal of Environmental Management, 251, 109569. http://dx.doi.org/10.1016/j.jenvman.2019.109569 PMid:31550603.
    » http://dx.doi.org/10.1016/j.jenvman.2019.109569
  • Somroo, A. A., Rehman, K. U., Zheng, L., Cai, M., Xiao, X., Hu, S., Mathys, A., Gold, M., Yu, Z., & Zhang, J. (2019). Influence of lactobacillus buchneri on soybean curd residue co-conversion by black soldier fly larvae (hermetia illucens) for food and feedstock production. Waste Management, 86, 114-122. http://dx.doi.org/10.1016/j.wasman.2019.01.022 PMid:30902235.
    » http://dx.doi.org/10.1016/j.wasman.2019.01.022
  • Su’i, M., Sumaryati, E., Dwianggraeni, F., Utomo, Y., Anggraini, N., & Rofiqoh, N. L. (2021). Monoglyceride biosynthesis from coconut milk with lypase enzyme of sesame seed sprouts as biocatalyst. Food Science and Technology, 41(Suppl. 1), 328-333.
  • Sun, S. D., Guo, J. J., & Chen, X. W. (2021). Biodiesel preparation from Semen Abutili (Abutilon theophrasti Medic.) seed oil using low-cost liquid lipase Eversa® transform 2.0 as a catalyst. Industrial Crops & Products, 169, 113643.
  • Surendra, K. C., Olivier, R., Tomberlin, J. K., Jha, R., & Khanal, S. K. (2016). Bioconversion of organic wastes into biodiesel and animal feed via insect farming. Renewable Energy, 98, 197-202. http://dx.doi.org/10.1016/j.renene.2016.03.022
    » http://dx.doi.org/10.1016/j.renene.2016.03.022
  • Talukder, M., Das, P., Tan, S. F., & Jin, C. W. (2011). Enhanced enzymatic transesterification of palm oil to biodiesel. Biochemical Engineering Journal, 55(2), 119-122.
  • Tschirner, M., & Simon, A. (2015). Influence of different growing substrates and processing on the nutrient composition of black soldier fly larvae destined for animal feed. Journal of Insects as Food and Feed, 1(4), 249- 259. http://dx.doi.org/10.3920/JIFF2014.0008
    » http://dx.doi.org/10.3920/JIFF2014.0008
  • Wang, C. W., Qian, L., Wang, W. G., Wang, T. L., Deng, Z. K., Yang, F., Xiong, J., & Feng, W. L. (2017a). Exploring the potential of lipids from black soldier fly: new paradigm for biodiesel production (I). Renewable Energy, 111, 749-756. http://dx.doi.org/10.1016/j.renene.2017.04.063
    » http://dx.doi.org/10.1016/j.renene.2017.04.063
  • Wang, X. M., Qin, X., Li, D. M., Yang, B., & Wang, Y. H. (2017). One-step synthesis of high-yield biodiesel from waste cooking oils by a novel and highly methanol-tolerant immobilized lipase. Bioresource Technology, 235, 18-24. http://dx.doi.org/10.1016/j.biortech.2017.03.086 PMid:28351728.
    » http://dx.doi.org/10.1016/j.biortech.2017.03.086
  • Wang, Y. H., Mai, Q. Y., Qin, X. L., Yang, B., Wang, Z. L., & Chen, H. T. (2010). Establishment of an evaluation model for human milk fat substitutes. Journal of Agricultural and Food Chemistry, 58(1), 642-649. http://dx.doi.org/10.1021/jf903048p PMid:20000702.
    » http://dx.doi.org/10.1021/jf903048p
  • Xu, T. T, Liu, L., Hou, S. L., Xu, J. X., Yang, B., Wang, Y. H., & Liu, J. S. (2012). Crystal structure of a mono- and diacylglycerol lipase from Malassezia globosa reveals a novel lid conformation and insights into the substrate specificity. Journal of Structural Biology, 178(3), 363-369. http://dx.doi.org/10.1016/j.jsb.2012.03.006 PMid:22484238.
    » http://dx.doi.org/10.1016/j.jsb.2012.03.006
  • Yin, Z. H., Zhu, L. D., Li, S. X., Hu, T. Y, Chu, R. Y, Mo, F., Hu, D., Liu, C. C, & Li, B. (2020). A comprehensive review on cultivation and harvesting of microalgae for biodiesel production: environmental pollution control and future directions. Bioresource Technology, 301, 122804. http://dx.doi.org/10.1016/j.biortech.2020.122804 PMid:31982297.
    » http://dx.doi.org/10.1016/j.biortech.2020.122804
  • Zheng, P. Y., Xu, Y., Wang, W. F., Qin, X. L., Ning, Z. X., Wang, Y. H., & Yang, B. (2014). Production of diacylglycerol-mixture of regioisomers with high purity by two-step enzymatic reactions combined with molecular distillation. Journal of the American Oil Chemists’ Society, 91(2), 251-259. http://dx.doi.org/10.1007/s11746-013-2365-2
    » http://dx.doi.org/10.1007/s11746-013-2365-2

Publication Dates

  • Publication in this collection
    10 Jan 2022
  • Date of issue
    2022

History

  • Received
    29 Sept 2021
  • Accepted
    02 Nov 2021
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