Bacillus sp.JY-23发酵制备胶原蛋白酶工艺优化  

Optimization of the Fermentation Process for Production of Collagenase by Bacillus sp.JY-23

作　　者：张荣 凌晓宁 邱露 李昆太 ZHANG Rong;LING Xiaoning;QIU Lu;LI Kuntai(College of Biosciences and Bioengineering/Jiangxi Engineering Laboratory for the Development and Utilization of Agricultural Microbial Resources,Jiangxi Agricultural University,Nanchang 330045,China;College of Food Science and Technology,Guangdong Ocean University,Zhanjiang,Guangdong 524088,China)

机构地区：[1]江西农业大学生物科学与工程学院/江西省农业微生物资源开发与利用工程实验室,江西南昌330045 [2]广东海洋大学食品科技学院,广东湛江524088

出　　处：《江西农业大学学报》2022年第5期1261-1271,共11页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(31760546)。

摘　　要：【目的】胶原蛋白具有特殊的三螺旋结构,对常见的蛋白酶具有抗性,只能被特定蛋白酶(胶原蛋白酶)水解。微生物胶原蛋白酶由于其较低的要求和较高的生产率,在胶原废物的高值化利用中发挥着重要作用。本研究拟通过单因素优化及响应面法优化,提高胶原蛋白酶产生菌的产酶量。【方法】通过单因素试验探究培养时间、温度、pH、接种量、装液量、碳源、氮源及金属离子等对胶原蛋白酶产生菌Bacillus sp.JY-23的产酶影响;在此基础上利用PB试验筛选出显著影响因素,最后通过CCD试验确定最佳发酵产酶工艺条件。【结果】通过PB试验筛选出葡萄糖、酵母浸粉、接种量和Fe^(2+)等4个显著影响因素,通过响应面法确定了最佳产酶发酵工艺:培养时间36 h、培养温度37℃、培养基pH值8.0、接种量2.97%,葡萄糖19.60 g/L、酵母浸粉25.80 g/L、Fe^(2+)0.75 mmol/L、NaCl 10 g/L。在该优化条件下,Bacillus sp.JY-23所产胶原蛋白酶活力达到28.13 U/mL,较优化前的酶活(5.94 U/mL)提高4.73倍。【结论】经过单因素、PB设计和响应面逐级优化,显著提高了Bacillus sp.JY-23发酵产胶原蛋白酶的酶活,为该菌株工业化应用于胶原蛋白活性肽开发提供了理论基础。[Objective]Collagen is resistant to common proteases due to its special triple helix structure,resulting in its hydrolysis by a specific protease(collagenase).Microbial collagenases play an important role in the recovery and high-value utilization of collagen waste because of its low requirements and high productivity.Therefore,this study intends to improve the enzyme production of collagenase producing bacteria by single factor optimization and response surface methodology.[Method]The effects of culture time,temperature,pH,inoculation concentration,loading capacity,carbon source,nitrogen source and metal ions on the enzyme production of collagenase producing strain Bacillus sp.JY-23 were studied by single factor test.Based on the test,the significant influencing factors were screened by PB test.Finally,the optimal fermentation conditions were determined by CCD test.[Result]Four significant influencing factors(glucose,yeast extract,inoculation amount and Fe^(2+))were identified by PB test.In addition,the optimum process conditions,culture cycle 36 h,culture temperature 37 ℃,pH 8.0,inoculation concentration 2.97%,glucose 19.60 g/L,yeast extract 25.80 g/L,Fe^(2+) 0.75 mmol/L and NaCl 10 g/L,were determined by response surface methodology.Under the optimized conditions,the collagenase activity of Bacillus sp.JY-23 reached 28.13 U/mL,which was 4.73 times higher than that before optimization(5.94 U/ml).[Conclusion]Through single factor,PB design and response surface step-by-step optimization,the collagenase activity produced by Bacillus sp.JY-23 fermentation is significantly improved,which provides a theoretical basis for the industrialized application of the strain in the development of collagen active peptides.

关 键 词：胶原蛋白 Bacillus sp.JY-23 Plackett-Burman试验设计 响应面法 

分 类 号：TS201.3[轻工技术与工程—食品科学]