重组耐热β-葡萄糖苷酶的固定化及其对黄酮糖苷的定向转化

doi:10.3969/j.issn.1673-5854.2021.03.002

Abstract

Abstract:

In order to increase the efficiency of the enzymatic conversion of flavonol glycosides and reduce the cost of the enzyme, the immobilization conditions of recombinant thermostable β-glucosidase were investigated. The differences of enzymatic properties between immobilized enzyme and free enzyme were compared, and the conversion curves and conversion efficiency of the immobilized enzyme on transforming three kinds of flavonol glycosides were obtained. The results showed that the optimal immobilization conditions were sodium alginate 1.5%, calcium chloride 2%, glutaraldehyde 1.25%, and pellets solidifying time 2.5 h under room temperature and pH 6.0. Under these conditions, enzyme activity was 0.64 U/mg, and the immobilization rate was 62.5%. The pH stability and temperature stability of the immobilized β-glucosidase were better than those of the free β-glucosidase and it retained more than 55% residual activity after reusing for 20 times. The isoquercetin, daidzin, and icariside I were transformed into their aglycones with high efficiency though immobilized β-glucosidase and the conversion rates were 99.37%, 97.21% and 97.93%, respectively, and the yields of quercetin, daidzein and icariin were 0.776, 1.091 and 0.714 g/(L·h), respectively.

Key words: flavonol glycosides, thermostable β-glucosidase, immobilization of enzyme

CLC Number:

TQ35

Jingcong XIE, Hao XU, Ning ZHANG, Jing YANG, Jian ZHAO. Immobilization of Thermostable Recombinant β-Glucosidase and Its Application in Transformation of Flavonol Glycosides[J]. Biomass Chemical Engineering, 2021, 55(3): 10-16.

Figures/Tables 6

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固定化条件 conditions		固定化酶活力/(U·mg^-1) activity of immobilized enzyme	酶固定化率/% rate of enzyme recovery
海藻酸钠质量分数 mass fraction of sodium alginate	1.0%	0.54	35.0±2.6
	1.5%	0.55	54.0±2.5
	2.0%	0.35	44.4±2.1
	2.5%	0.21	32.3±1.8
	3.0%	0.16	29.8±1.2
	3.5%	0.13	27.1±2.9
氯化钙质量分数 mass fraction of calcium chloride	1.0%	0.33	31.3±2.6
	1.5%	0.46	45.0±3.2
	2.0%	0.57	55.0±2.8
	2.5%	0.60	56.6±2.7
	3.0%	0.44	43.0±2.3
	3.5%	0.42	39.3±2.0
颗粒硬化时间 pelletssolidifying time	1.0 h	0.26	23.0±1.6
	1.5 h	0.35	35.4±2.1
	2.0 h	0.47	43.4±1.3
	2.5 h	0.56	55.7±2.7
	3.0 h	0.59	55.4±2.3
	3.5 h	0.53	54.4±1.3
	4.0 h	0.56	53.4±2.3
戊二醛质量分数 mass fraction of glutaraldehyde	0%	0.32	30.1±2.3
	0.5%	0.44	43.7±3.6
	1.0%	0.57	53.5±2.3
	1.25%	0.64	62.5±2.4
	1.5%	0.60	57.7±1.6
	2.0%	0.59	55.3±3.1

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Immobilization of Thermostable Recombinant β-Glucosidase and Its Application in Transformation of Flavonol Glycosides

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