Abstract The present study aimed to identify the bioactive constituents in the chloroform extract of H. spicatum rhizomes (HS-RCLE), further evaluated for its in-vitro pesticidal activities validating via molecular docking techniques. GC/MS analysis of HS-RCLE identified 14 compounds contributing 84.1 % of the total composition. The extract was dominated by oxygenated sesquiterpenes (43.1 %) with curcumenone (25.2 %) and coronarin E (14.8 %) as the major compounds. The extract recorded 89.4 % egg hatchability inhibition and 82.6 % immobility of Meloidogyne incognita, 66.7 % insecticidal activity on Spodoptera litura, 100 % phytotoxic activity on Raphanus raphanistrum seeds, and 74.7 % anti-fungal activity on Curvularia lunata at the respective highest dose studied. The biological activities were furthermore validated by using docking studies on certain proteins/enzymes namely acetylcholinesterase (PBD ID: IC2O), carboxylesterase (PDB ID: 1CI8), acetohydroxyacid synthase (PBD ID: 1YHZ) and trihydroxy naphthalene reductase (PBD ID: 3HNR). The bioactivity of the major constituents of the extract was predicted with the help of in silico PASS studies. HS-RCLE was observed to be a viable alternative source of natural pesticidal agents and paves the way for further studies on its mechanistic approaches and field trials to ascertain its pesticidal studies.

Intraductal papillary neoplasm of the bile duct (IPN-B) is a rare preinvasive intraductal pathology of the biliary tract. It should be differentiated from other more common benign or malignant causes of biliary obstruction and dilatation such as calculi or cholangiocarcinoma because the management and prognosis of this condition differs significantly. This case report describes a case of IPN-B in a 45-year-old female patient who presented with non-specific complaints of chronic abdominal pain without jaundice for three months.

Poly-ß-hydroxybutyrate (PHB) is a biodegradable intracellular microbial product produced by many bacteria and it is comparable to some of the petrochemical derived thermoplastics such as polypropylene. One of the main barriers for the commercial exploitation is the high cost of the substrate for the production of biopolymer. The utilization of mixed microbial cultures facilitates the use of complex substrates thereby reducing the cost of PHB production. In the present study, mixed culture systems were evaluated for PHB production. Bacillus firmus NII 0830 was used for the production of PHB since it accumulates a large amount of PHB and a second organism Lactobacillus delbrueckii NII 0925 was used to provide lactic acid. FTIR and 1H NMR analyses revealed that the PHB extracted from pure culture and mixed culture showed exact match to that of standard PHB. Biodegradation studies of the PHB blends showed 87% degradation. It was also found that a consortium of organisms degraded the films faster than a single organism.

A bioprocess was developed for extracellular MAP production from Streptomyces gedanensis by solid-state fermentation. Response surface methodology of Box Behken Design was performed to evaluate the interaction effects of most significant variables {inoculum size, (NH4)2SO4 concentration, MgSO4.7H2O and tryptone) on MAP production after the single parameter optimization and it resulted a maximum MAP production of 55.26 IU/g PUF after 120 h of fermentation. The concentrated crude MAP displayed a pH and temperature optimum of 8.5 and 50°C. By analyzing the thermal stability, the MAP was found to be stable in a temperature range of 50 to 55°C but lost about 50% of its activity at 65°C after 30 min. This is a first report of this kind of study for MAP.

Since centuries vegetable oils are consumed as human food but it also finds applications in biodiesel production which is attracting more attention. But due to being in competition with food it could not be sustainable and leads the need to search for alternative. Nowdays microbes-derived oils (single cell oils) seem to be alternatives for biodiesel production due to their similar composition to that of vegetable oils. However, the cold flow properties of the biodiesel produced from microbial oils are unacceptable and have to be modified by an efficient transesterification. Glycerol which is by product of transesterification can be valorised into some more useful products so that it can also be utilised along with biodiesel to simplify the downstream processing. The review paper discusses about various potent microorganisms for biodiesel production, enzymes involved in the lipid accumulation, lipid quantification methods, catalysts used in transesterification (including enzymatic catalyst) and valorisation of glycerol.

The aim of this work was to study the production of poly-3-hydroxybutyrate (PHB) under nitrogen limited conditions by Bacillus sphaericus NII 0838 using crude glycerol from biodiesel industry as sole carbon source. Effect of various process parameters on PHB production such as glycerol concentration, inoculum size and pH of the medium were optimized. Characterization of extracted PHB was carried out by FT-IR, ¹H and 13C NMR. Results showed that the bacterial culture accumulated about 31% PHB in crude glycerol medium. The extracted PHB was blended with other polymers to improve its physical characteristics. The thermal properties of the polymer like melting temperature (Tm) and heat of fusion (ΔHf) were determined using DSC.

The proteolytic system of lactic acid bacteria contribute to the development of flavor during the ripening of cheese through the generation of short peptides and free amino acids, which directly or indirectly act as flavor precursors. Newly isolated lactic acid bacteria (LAB) as well as those procured from culture collection centers were screened for the production of various substrate specific aminopeptidases. Among all the strains screened, L. brevis (NRRL B-1836) was found to produce quantifiable amount of intracellular arginine specific aminopeptidase (EC 3.4.11.6). The productivity of arginine aminopeptidase in 5 L fermentor was 36 IU/L/h. The Luedeking and Piret model was tested for intracellular production of aminopeptidase and the data seemed to fit well, as the correlation coefficient was 0.9964 for MRS. The αAP and βAP was 0.4865 and 0.0046, respectively in MRS medium indicating that the yield was predominantly depended on growth. The culture produced lactic acid and also tolerated pH 2.0-3.0 and 0.3-0.5% bile salts, the most important probiotic features.

O ácido giberélico (GA3) é um importante hormônio vegetal. A fermentação no estado sólido (FES) utiliza resíduos agro-industriais reduzindo os custos de produção. Neste trabalho a seleção de cepas (quatro de Gibberella fujikuoroi e uma de Fusarium moniliforme) e substratos (polpa cítrica, casca de soja, bagaço de cana, farelo de soja, bagaço de mandioca e casca de café) e o estudo da preparação do inóculo foram conduzidos para otimizar as condições de produção de GA3 por FES. Os ensaios foram realizados em frascos de erlenmeyer a 29°C, com umidade inicial de 75-80%. Diferentes meios para a produção do inóculo foram testados em relação à viabilidade das células e produção de GA3 por FES. F. moniliforme LPB03 e polpa cítrica foram escolhidos. O melhor meio para a produção de inóculo foi o extrato de polpa cítrica. A produção por FES alcançou 5.8 g de GA3/kg de polpa cítrica após 3 dias de fermentação.

Gibberellic acid (GA3) is an important hormone, which controls plant's growth and development. Solid State Fermentation (SSF) allows the use of agro-industrial residues reducing the production costs. The screening of strains (four of Gibberella fujikuoroi and one of Fusarium moniliforme) and substrates (citric pulp, soy bran, sugarcane bagasse, soy husk, cassava bagasse and coffee husk) and inoculum preparation study were conducted in order to evaluate the best conditions to produce GA3 by SSF. Fermentation assays were carried out in erlenmeyers flasks at 29°C, with initial moisture of 75-80%. Different medium for inoculum production were tested in relation to cells viability and GA3 production by SSF. F. moniliforme LPB 03 and citric pulp were chosen for GA3 production. The best medium for inoculum production was citric pulp extract supplemented with sucrose. GA3 production by SSF reached 5.9 g /kg of dry CP after 3 days of fermentation.

The aim of this work was to study the production of polyhydroxybutyrate (PHB) using agro- industrial residues as the carbon source. Seven substrates, viz., wheat bran, potato starch, sesame oil cake, groundnut oil cake, cassava powder, jackfruit seed powder and corn flour were hydrolyzed using commercial enzymes and the hydrolyzates assessed for selecting the best substrate for PHB production. Jackfruit seed powder gave the maximum production of PHB under submerged fermentation using Bacillus sphaericus (19%) at the initial pH of 7.5.

As propriedades das resinas de troca iônica, da Amberlite IRA 402, uma resina de troca aniônica forte, e da IRA 67, uma resina de troca aniônica fraca, foram determinadas para se avaliar a adequabilidade comparativa delas à obtenção de ácido lático de bagaço de mandioca fermentado. Dados sobre a capacidade de ligação e sobre a obtenção provaram que a resina de base fraca na forma de cloreto era a mais adequada para a obtenção de ácido lático em soluções aquosas e meios de fermentação. Os meios de fermentação obtidos da sacarificação e da fermentação simultâneas de meios baseados hidrolisados de fécula de bagaço de mandioca foram usados para o estudo da obtenção de ácido lático usando uma coluna de resina de base fraca. A Amberlite IRA 67 mostrou-se muito mais eficaz do que a Amberlite IRA 402 para a obtenção de ácido lático. Como em outros relatórios, devido à presença de nutrientes e íons que não lactatos, a capacidade de ligação foi ligeiramente inferior enquanto se utilizavam meios fermentados em vez de soluções ácidas láticas aquosas.

The properties of the ion exchange resins, Amberlite IRA 402, a strong anion exchange resin and IRA 67, a weak anion exchange resin were determined to evaluate their comparative suitability for lactic acid recovery from fermented cassava bagasse. Data on binding capacities and recovery proved that weak base resin in chloride form was the most favourable ones for lactic acid recovery from aqueous solutions and fermentation media. Fermented media obtained through simultaneous saccharification and fermentation of cassava bagasse starch hydrolysate based medium were used for lactic acid recovery study using weak base resin column. Amberlite IRA 67 had much more efficiency than Amberlite IRA 402 to recover lactic acid. Like in other reports, due to the presence of nutrients and ions other than lactate, the binding capacity was slightly lesser while using fermented media (~93%) instead of aqueous lactic acid solutions (~98%).

O objetivo do presente trabalho foi estudar o efeito da cafeína e taninos sobre o desenvolvimento do fungo Pleurotus sp. e avaliar a possibilidade do uso da casca de café como substrato no seu cultivo. Oito cepas de P. ostreatus e duas cepas de P. sajor-caju foram selecionadas em um meio à base de extrato aquoso de casca de café e Agar. A cepa P. ostreatus LPB 09 foi escolhida para estudos posteriores com base na velocidade de crescimento do micélio e produção de biomassa. O estudo do desenvolvimento do cogumelo em meio com diferentes concentrações de cafeína mostrou que o aumento da concentração da mesma influencia negativamente crescimento. A partir de uma concentração de cafeína de 2500 mg/L não foi observado crescimento. Observou-se ainda que o Pleurotus não degradou a cafeína, mas a absorveu. Os taninos em concentração abaixo de 100 mg/L no meio estimularam o crescimento do micélio, no entanto acima de 500 mg/L mostraram efeito negativo. Quando a concentração de taninos atingiu 1000 mg/L, o cogumelo ainda sobreviveu e mostrou uma certa tolerância. O ácido tânico não foi encontrado no micélio, o que confirma que o Pleurotus possui a capacidade de degradá-lo. A frutificação ocorreu após 20 dias de inoculação e a eficiência biológica atingiu 97% em 60 dias. O conteúdo de cafeína da casca de café foi reduzido para 60.7% e o de taninos para 79.2%. Os resultados mostram boas perspectivas para o cultivo de Pleurotus utilizando casca de café como substrato sem a necessidade de qualquer pré-tratamento.

The objective of this work was to study the effect of caffeine and tannins on Pleurotus sp. cultivation and to evaluate the feasibility of using coffee husks as substrate for mushroom cultivation. Eight strains of P. ostreatus and two strains of P. sajor-caju were screened on a medium prepared from agar extract of coffee husk. Based on best mycelial growth and biomass production, the strain P. ostreatus LPB 09 was selected for detailed studies. With the increase of caffeine concentration, the mycelial growth and the biomass production decreased, and no growth was observed when concentration of caffeine was 2500 mg/L. Furthermore, Pleurotus did not degrade the caffeine, but absorbed it. Tannin under 100 mg/L in the medium stimulated the growth of mycelia, but above 500 mg/L it had a negative effect. When the concentration reached 1000 mg/L, the fungus still survived and showed a certain tolerance to it. No tannic acid was found in the mycelia, but its concentration decreased in the medium. This fact confirmed that Pleurotus had the capacity of degrading tannic acid. Fructification occurred after 20 days of inoculation and the biological efficiency reached about 97% after 60 days. Caffeine content in the husk after cultivation was reduced to 60.7% and tannins to 79.2%. The results indicated the feasibility of using coffee husk without any pretreatment for the cultivation of Pleurotus.

Farelo de arroz foi utilizado como substrato para seleção de nove linhagens de Rhizopus sp. com vistas a produção de protease neutra. A linhagem que apresentou maior produtividade da enzima foi Rhizopus microsporus NRRL 3671, sendo utilizada na otimização dos parâmetros do processos e produção da enzima. As condições otimizadas para produção da enzima foram 44% de umidade inicial, temperatura de 30ºC e 72h de fermentação.A suplementação do farelo de arroz com uma fonte de carbono favoreceu a produção da enzima, porém a adição de caseína resultou em um aumento marginal do rendimento em protease. Condições otimizadas foram utilizadas para obtenção do extrato cru da enzima que foi parcialmente purificado por precipitação e diálise. A enzima purificada teve sua atividade incrementada 3 vezes. A enzima foi classificada como metalo-protease, sendo ativada pelo Mn2+ , sendo que sua atividade máxima foi obtida a temperatura de 60ºC e a pH 7.0.

Rice bran was used as the substrate for screening nine strains of Rhizopus sp. for neutral protease production by solid-state fermentation. The best producer, Rhizopus microsporus NRRL 3671, was used for optimizing the process parameters for enzyme production. Fermentation carried out with 44.44 % initial moisture content at a temperature of 30 C for 72 h was found to be the optimum for enzyme secretion by the fermenting organism. While most of the carbon supplements favored enzyme production, addition of casein resulted in a marginal increase in protease yield. Fermentation was then carried out under optimized conditions to obtain the crude extract of the enzyme, which was partially purified by precipitation and dialysis. A 3-fold increase in the enzyme purity was achieved in this manner. The enzyme was found to be a metalloprotease, being activated by Mn2+, with maximal activity at a temperature of 60 C and pH 7.0.

Fungos do gênero Monascus são uma fonte promissora de corantes vermelhos naturais. No entanto, antes de se aplicar Monascus a alimentos, é importante selecionar linhagens capazes de produzir grandes quantidades de pigmentos, com o mínimo possível de citrinina. Além disso, a estabilidade de cor desses pigmentos deve ser adequadamente investigada. Com o objetivo de comparar linhagens de Monascus para a produção de biopigmentos em fermentação em substrato sólido (FSS), 4 linhagens (NRRL 1991, NRRL 2897, CCT 3802 e LPB 31) foram cultivadas sobre PDA em placas de Petri, e comparadas com relação à velocidade de crescimento radial. Além disso, essas linhagens foram cultivadas em arroz cozido, e comparadas quanto à sua capacidade de produção de biopigmentos e de citrinina. Os resultados mostram que a linhagem LPB 31 é a melhor linhagem para produção de biopigmentos em FSS, dando tanto maior concentração de pigmentos quanto menor concentração de citrinina, mostrando que se trata de uma linhagem promissora para a obtenção deste bioproduto. Os biopigmentos extraídos dos substratos fermentados foram testados quanto à estabilidade frente ao calor e ao pH, mostrando que os biopigmentos de Monascus são instáveis em pH's baixos e altas temperaturas, mas que podem ser usados sem problema em condições de neutralidade, em alimentos sem processamento térmico.

Fungi form the genus Monascus are a promising source for natural color additives. However, before effectively applying Monascus to foods, it is important to select strains which produce large amounts of biopigments but little or no citrinin, a mycotoxin usually also produced by these fungi. Also, color stability of these pigments should be properly investigated. In order to compare Monascus strains for biopigment production in solid substrate fermentation (SSF), 4 strains (NRRL 1991, NRRL 2897, CCT 3802 and LPB 31) were cultivated over PDA in Petri dishes, and compared for radial growth velocity. Also, these strains were cultivated over cooked rice, and compared in relation to their capacity to produce biopigments and citrinin. The results showed that the strain LPB 31 is the best strain for biopigment production in SSF, giving both higher pigment concentration and lower citrinin concentration on the extracts, showing that it is a promising strain for production of this bioproduct. Biopigmentassays for heat and pH stability, show that these biopigments are unstable at low pH and high temperatures, but may be successfully used at near-neutrality pH's and in non-thermal processed foods.

Fermentação no Estado Sólido foi empregada na produção de alfa-amilase usando Aspergillus niger. Diferentes tipos de torta foram utilizadas, como torta de óleo de coco (COC), torta de de óleo de amendoim (GOC) torta de óleo de sesamo (SOC), torta de palma (PKC) e torta de óleo de oliva (OOC) foram selecionadas para serem usadas como substratos para produção de enzima e comparadas com o farelo de trigo (WB), GOC foi escolhido por ser o que produziu maiores concentrações de enzima. A combinação WB e GOC (1:1) resultou em maiores títulos da enzima quando em comparação com os substratos individuais. A máxima concentração de enzima (9196 U/ gms) foi obtida quando a FES foi conduzida utilizando WB + GOC, com umidade de 64% e suplementada com lactose e nitrato de amônia (1% cada) a 300C por 72 horas utilizando 2 mL de uma suspensão de esporo (6x107sporos/ml). A purificação parcial da enzima usando frações de sulfato de amônio resultou num aumento de 2-4 vezes o aumento da atividade. A enzima apresentou um peso molecular de 68 Kda pelo SDS_PAGE. Exceto Mn, todos os outros íons metálicos como Ca, K, Na, Mg são inibitórios na produção da enzima.

Solid-state fermentation was carried out for the production of alpha-amylase using Aspergillus oryzae. Different oil cakes such as coconut oil cake (COC) sesame oil cake (SOC), groundnut oil cake (GOC), palm kernel cake (PKC) and olive oil cake (OOC) were screened to be used as substrate for the enzyme production and also compared with wheat bran (WB). GOC was found to be the best producer of the enzyme among these. Combination of WB and GOC (1:1) resulted higher enzyme titres than the individual substrates. Maximum amount of enzyme (9196 U/gds) was obtained when SSF was carried out using WB + GOC, having initial moisture of 64% and supplemented with lactose and ammonium nitrate (1% each) at 30ºC for 72h using 2 mL spore suspension (6x10(7)spores/ml). Partial purification of the enzyme using ammonium sulphate fractionation resulted in 2.4-fold increase in the activity. The enzyme showed molecular weight of 68 KDa by SDS-PAGE. Except Mn, all other metal ions such as Ca, K, Na, Mg were found to be inhibitory for the enzyme activity. The enzyme was optimally active at 50(0)C and pH 5.0.