枯草芽孢杆菌WB600角蛋白酶重组菌发酵条件优化

doi:10.16431/j.cnki.1671-7236.2020.01.006

Abstract

Abstract: The aim of this experiment was to optimize the fermentation conditions of the constructed keratinase recombinant B.subtilis WB600 strain WB600-K,and obtain the optimal enzyme production conditions in order to increase the yield of recombinant keratinase;At the same time,the combination optimization between different types of carbon sources and different nitrogen sources was studied in order to obtain a low-cost recombinant fermentative medium formula.The results showed that when the fermentation D_{600 nm}=1.0,and 0.5% xylose was added to induce fermentation,the expression of keratinase was best in the recombinant WB600-K (P<0.05).As a carbon source,sorbitol could significantly increase the activity of the recombinant strain WB600-K keratinase (P<0.05),but the cost was too high to be a source of mixed carbon sources.The combination of different carbon sources (glucose,corn flour,molasses and glycerin) could significantly increase the activity of keratinase (P<0.05).The combination of different kinds of nitrogen sources (peptone,yeast powder,soybean meal and urea) significantly increased the activity of keratinase less than single nitrogen source (peptone) (P<0.05).The WB600-K had the highest keratinase activity at 48 h after induction of fermentation (P<0.05).The overall experimental optimization results showed that,at the condition of corn flour 5.1 g/L,glucose 5.9 g/L,molasses 5.9 g/L,glycerol 3.0 g/L,peptone 20 g/L,NaCl 10 g/L,0.5% xylose was added when fermentation D_{600 nm}=1.0,and 48 h fermentation,the activity of keratinase could be increased significantly (P<0.05),which was 56.9 U/mL and 49.74% higher than that in initial culture conditions.In summary,fermentation under optimized fermentation conditions could increase the activity of recombinant strain WB600-K keratinase,which laid a foundation for the industrial production of keratinase.

Key words: B.subtilis WB600; keratinase; fermentative conditions; optimization

CLC Number:

S616.3

LIAO Chaoyong, ZHANG Tieying, LIU Junli, SUI Jingwei. Optimization of the Fermentation Conditions of Recombinant B.subtilis WB600 Produced Keratinase[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 44-52.

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Optimization of the Fermentation Conditions of Recombinant B.subtilis WB600 Produced Keratinase

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