Treatment of sugarcane bagasse for the immobilization of soybean β-glucosidase and application in soymilk isoflavones

Moreira, Amanda Aleixo; Ferreira, Ana Camila Vaitkevicius; Silva, Josemeyre Bonifácio da; Ribeiro, Mara Lúcia Luiz

doi:10.1590/1981-6723.24318

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Brazilian Journal of Food Technology

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ORIGINAL ARTICLE • Braz. J. Food Technol. 22 • 2019 • https://doi.org/10.1590/1981-6723.24318 copy

Treatment of sugarcane bagasse for the immobilization of soybean β-glucosidase and application in soymilk isoflavones

Tratamento de bagaço de cana-de-açúcar para imobilização de β-glucosidase de soja e aplicação em isoflavonas de soja

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Sugarcane bagasse from agro-industrial residues was autoclaved at 121 °C for 15 min, treated with 2% NaOH and activated with 2.5% glutaraldehyde for the immobilization of soybean β-glucosidase. Scanning electron microscopy, energy dispersive X-ray spectroscopy and Fourier transform infrared spectroscopy characterized and confirmed the immobilization of the β-glucosidase on the sugarcane bagasse. The immobilization efficiency was influenced by the type of bagasse modification and was 99% at maximum. The optimum immobilization conditions were 1 mg mL^-1 protein, pH 7.0, 2.5% glutaraldehyde, 110 rpm and 8 h of incubation at 4 °C. The immobilized system could be reused for 15 cycles without the complete loss of activity. The thermal stability indicated a residual activity of 15% after 180 min incubation at 70 °C. The conversion efficiency of glucosides to aglycones in commercial soymilk by β-glucosidase immobilized on sugarcane bagasse was evaluated and the total aglycone content increased by 23.8% after incubation at 50 °C for 120 min.

Keywords:
Soybean; β-glucosidase; Sugarcane bagasse; Enzyme immobilization; Isoflavones; Commercial soymilk

Modifications	Protein (0.7 mg mL^-1)	Protein (1.4 mg mL^-1)
Modifications	IE (%)	IE (%)
SCB_{1(Water 60 °C)}	66 ± 2 ^Ac*	62 ± 4 ^Ac
SCB_{2(Water 60 °C + 2.5% glutaraldehyde)}	77 ± 3 ^Ab	80 ± 1 ^Ab
SCB_{3(2% NaOH + autoclave)}	79 ± 2 ^Ab	81 ± 2 ^Ab
SCB_{4(2% NaOH + autoclave + 2.5% glutaraldehyde)}	98 ± 1^Aa	97 ± 1 ^Aa
SCB_{5(70% Ethanol + autoclave)}	56 ± 3^Ad	53 ± 3 ^Ad
SCB_{6(70% Ethanol + autoclave +2.5% glutaraldehyde)}	82 ±1 ^Ab	83 ± 1^Ab

Treatment	Cellulose (%)	Lignin (%)	Hemicellulose (%)	Ash (%)
Bagasse _(washed)	45.6 ± 3 ^C*	5.6 ± 1 ^A	42.6 ± 2 ^A	0.79 ± 0.1 ^A
Bagasse _{(2% NaOH)}	59.5 ± 2 ^B	1.9 ± 0.7 ^B	34.5 ± 2 ^B	0.66 ± 0.05 ^A
Bagasse _{(2% NaOH + autoclaved)}	74.0 ± 3 ^A	1.6 ± 0.4 ^B	21.6 ± 1 ^C	0.61 ± 0.06 ^A

Isoflavones (µmol mL^-1)	Control	Treatments
		min/25 °C			min/50 °C
		30	60	120	30	60	120
Daidzin	0.127 ± 0.002 ^A*	0.116 ± 0.001 ^B	0.115 ± 0.001 ^B	0.116 ± 0.001 ^B	0.116 ± 0.001 ^B	0.115 ± 0.001 ^B	0.116 ± 0.001 ^B
Genistin	0.184 ± 0.004 ^A	0.157 ± 0.004 ^B	0.154 ± 0.003 ^B	0.153 ± 0.004 ^B	0.155 ± 0.002 ^B	0.154 ± 0.001 ^B	0.154 ± 0.002 ^B
Malonyldaidzin	0.025 ± 0.001 ^A	0.025 ± 0.002 ^A	0.025 ± 0.001 ^A	0.026 ± 0.001 ^A	0.026 ± 0.001 ^A	0.026 ± 0.001 ^A	0.026 ± 0.00 ^A
Malonylgenistin	0.021 ± .001 ^A	0.018 ± 0.001 ^B	0.018 ± 0.001 ^B	0.017 ± 0.001 ^B	0.018 ± 0.002 ^B	0.018 ± 0.001 ^B	0.017 ± 0.002 ^B
Daidzein	ND	ND	ND	ND	ND	ND	ND
Genistein	0.021 ± 0.001 ^D	0.023 ± 0.0005 ^C	0.023 ± 0.0005 ^C	0.024 ± 0.0005 ^B	0.024 ± 0.0005 ^B	0.025 ± 0.0005 ^B	0.026 ± 0.0005^A
TOTAL	0.378 ± 0.005 ^A	0.339 ± 0.004 ^B	0.336 ± 0.004 ^B	0.337 ± 0.003 ^B	0.339 ± 0.004 ^B	0.338 ± 0.003 ^B	0.339 ± 0.002 ^B

Instituto de Tecnologia de Alimentos - ITAL Av. Brasil, 2880, 13070-178 Campinas - SP / Brasil, Tel 55 19 3743-1762 - Campinas - SP - Brazil
E-mail: bjftsec@ital.sp.gov.br

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[1] *Corresponding Author: Josemeyre Bonifácio da Silva, Universidade Estadual de Londrina (UEL), Departamento de Bioquímica e Biotecnologia, Caixa Postal: 6001, CEP: 86051-990, Londrina/PR - Brasil, e-mail: josibonifacio0576@gmail.com