一株瘤胃源产纤维素酶菌株的培养条件优化
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摘要
山羊瘤胃内容物菌群中分离得到的一株产纤维素酶菌株T15,采用3,5-二硝基水杨酸法(DNS法)进行优化。对菌株培养基成分碳源、氮源和金属离子进行优化时发现,6 g/L的葡萄糖为碳源和5 g/L的牛肉膏为氮源时,产生酶活力达到最高。培养基中添加0.03 mol/L的Ca~(2+),对菌株产生纤维素酶有促进作用。在最优发酵培养基成分为基础,对菌株T15的发酵培养温度、培养时间、培养pH条件和接种量等四因素、三水平进行正交试验,该菌株的产纤维素酶最优发酵条件:以9 mL为接种量,培养基pH值7,培养温度35℃和培养时间40 h,其产生酶活力达到26.76 U/mL;其中培养时间、接种量、培养温度、和培养基酸碱条件对菌株发酵产纤维素酶影响依次增大。
A cellulase-producing strain T15 was isolated from the rumen content flora of goats. The fermentation conditions of the strain were optimized by 3,5-Dinitrosalicylic acid(DNS) method. It was found that when 6 g/L glucose was carbon source and 5 g/L beef extract was nitrogen source, the strain had the best cellulase production characteristics and the highest enzyme activity. Adding 0.03 mol/L Ca~(2+) to the medium could obviously promote the enzyme production of the strain. Based on the optimum fermentation medium composition, orthogonal experiments were carried out on four factors and three levels of strain T15: fermentation temperature, culture time, culture pH condition and inoculation amount.The optimum fermentation conditions for cellulase production of strain T15 were as follows: 9 mL as inoculation amount,pH7, and 26.76 U/mL as enzyme activity. The incubation temperature was 35 ℃ and incubation time was 40 h. The effects of incubation time, inoculation amount, incubation temperature and medium acid-base conditions on cellulase production by strains increased in turn.
A cellulase-producing strain T15 was isolated from the rumen content flora of goats. The fermentation conditions of the strain were optimized by 3,5-Dinitrosalicylic acid(DNS) method. It was found that when 6 g/L glucose was carbon source and 5 g/L beef extract was nitrogen source, the strain had the best cellulase production characteristics and the highest enzyme activity. Adding 0.03 mol/L Ca~(2+) to the medium could obviously promote the enzyme production of the strain. Based on the optimum fermentation medium composition, orthogonal experiments were carried out on four factors and three levels of strain T15: fermentation temperature, culture time, culture pH condition and inoculation amount.The optimum fermentation conditions for cellulase production of strain T15 were as follows: 9 mL as inoculation amount,pH7, and 26.76 U/mL as enzyme activity. The incubation temperature was 35 ℃ and incubation time was 40 h. The effects of incubation time, inoculation amount, incubation temperature and medium acid-base conditions on cellulase production by strains increased in turn.
引文
[1]田丽萍.反刍动物生理代谢与瘤胃微生物关系的研究[J].黑龙江畜牧兽医,2016(8):49-51.
[2]马涛.瘤胃微生物多样性与定量[J].动物营养学报,2015,27(23):3649-3654.
[3]谢丽静,王伟华,王海宽,等.丁酸梭菌的应用及对食品安全的影响[J].中国食物与营养,2017(7):5-9.
[4]高培基.纤维素酶降解机制及纤维素酶分子结构与功能研究进展[J].自然科学进展,2003(13):21-29.
[5]易中华.饲用微生态制剂丁酸梭菌的研究与应用进展[J].饲料研究,2012(2):14-17.
[6]李蒙,马淑华,杨瑞生.用纤维素酶、酵母菌处理玉米秸饲喂育肥羊的试验[J].山东畜牧兽医,2002(5):2-3.
[7]李明华,张大伟.饲料纤维素酶的研究与应用进展[J].畜牧与饲料,2006(7):28-33
[8]张广臣,雷虹.微生物发酵培养基优化中的现代数学统计学方法[J].食品与发酵工业,2010,36(5):110-113.
[9]赵新刚.洗涤用碱性纤维素酶及其产生菌的分离方法[J].微生物学通报,1999,26(1):63-65.
[10]张璐.不同pH下瘤胃液中纤维素降解产氢菌群的性能[J].大连工业大学学报,2017(5):328-332.
[11]吕东海,周岩民.饲料纤维素酶应用研究[J].中国饲料,2004(9):25-27.
[12]费尚芬,鹿宁.白腐菌纤维素酶高酶活菌株的筛选[J].安徽农业科学,2006,34(1):22-23.
[13]赵苗苗,玉柱.添加乳酸菌及纤维素酶对象草青贮品质的改善效果,2015,23(1):205-209.
[2]马涛.瘤胃微生物多样性与定量[J].动物营养学报,2015,27(23):3649-3654.
[3]谢丽静,王伟华,王海宽,等.丁酸梭菌的应用及对食品安全的影响[J].中国食物与营养,2017(7):5-9.
[4]高培基.纤维素酶降解机制及纤维素酶分子结构与功能研究进展[J].自然科学进展,2003(13):21-29.
[5]易中华.饲用微生态制剂丁酸梭菌的研究与应用进展[J].饲料研究,2012(2):14-17.
[6]李蒙,马淑华,杨瑞生.用纤维素酶、酵母菌处理玉米秸饲喂育肥羊的试验[J].山东畜牧兽医,2002(5):2-3.
[7]李明华,张大伟.饲料纤维素酶的研究与应用进展[J].畜牧与饲料,2006(7):28-33
[8]张广臣,雷虹.微生物发酵培养基优化中的现代数学统计学方法[J].食品与发酵工业,2010,36(5):110-113.
[9]赵新刚.洗涤用碱性纤维素酶及其产生菌的分离方法[J].微生物学通报,1999,26(1):63-65.
[10]张璐.不同pH下瘤胃液中纤维素降解产氢菌群的性能[J].大连工业大学学报,2017(5):328-332.
[11]吕东海,周岩民.饲料纤维素酶应用研究[J].中国饲料,2004(9):25-27.
[12]费尚芬,鹿宁.白腐菌纤维素酶高酶活菌株的筛选[J].安徽农业科学,2006,34(1):22-23.
[13]赵苗苗,玉柱.添加乳酸菌及纤维素酶对象草青贮品质的改善效果,2015,23(1):205-209.