灰色链霉菌LD1810产胰蛋白酶发酵条件的优化
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摘要
胰蛋白酶(Trypsin)作用于碱性氨基酸精氨酸及赖氨酸羧基所组成的肽键,被广泛用于食品、医药和皮革等工业领域。该文研究高产诱变灰色链霉菌LD1810产胰蛋白酶(SGT)的最优发酵培养基,探讨SGT的最佳发酵条件,为SGT的商业化生产提供技术参考。通过单因素试验和L_9(3~4)正交试验确定发酵培养基的最佳组分:葡萄糖20 g/L,大豆粉40 g/L,KH_2PO_4 2.5 g/L,CaCl_2 1 g/L。最适发酵条件为发酵温度28℃,初始pH 8.0,300 mL三角瓶的装液量100 mL,摇床转速240 r/min。
Trypsin acts on peptide bonds formed by carboxyl groups of basic amino acids arginine and lysine, and widely used in food, medicine and leather industries. The aim of this study was to investigate the optimum fermentation medium of SGT produced by Streptomyces griseus LD1810, the optimum fermentation conditions of SGT was explored to provide technical support for the commercial production of SGT. Single factor test and L_9(3~4) orthogonal design were used to determine the optimal composition of the fermentation mediuml as follows: glucose 20 g/L, soybean cake 40 g/L, KH_2PO_4 2.5 g/L,CaCl_2 1 g/L. The optimal fermentation conditions obtained were as follows: culture temperature of 28 ℃, initial pH of 8.0, 100 mL broth content in 300 mL triangle bottle and rotating speed 240 r/min.
Trypsin acts on peptide bonds formed by carboxyl groups of basic amino acids arginine and lysine, and widely used in food, medicine and leather industries. The aim of this study was to investigate the optimum fermentation medium of SGT produced by Streptomyces griseus LD1810, the optimum fermentation conditions of SGT was explored to provide technical support for the commercial production of SGT. Single factor test and L_9(3~4) orthogonal design were used to determine the optimal composition of the fermentation mediuml as follows: glucose 20 g/L, soybean cake 40 g/L, KH_2PO_4 2.5 g/L,CaCl_2 1 g/L. The optimal fermentation conditions obtained were as follows: culture temperature of 28 ℃, initial pH of 8.0, 100 mL broth content in 300 mL triangle bottle and rotating speed 240 r/min.
引文
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[2]AMINO R,TANAKA A S,SCHENKMAN S.Triapsin,an unusual activatable serine protease from the saliva of the hematophagous vector of Chagas’disease Triatoma infestans(Hemiptera:Reduviidae)[J].Insect Biochem Mol Biol,2001,31(4/5):465-472.
[3]黄康健.胰蛋白酶治疗蛇咬伤336例报告[J].中国医刊,1979(5):33.
[4]孙逸佃.胰蛋白酶治疗蜈蚣咬伤[J].中国医院药学杂志,1985,5(2):37-38.
[5]刘欣华,刘奕志,夏朝霞.胰蛋白酶预防后发性白内障的实验研究[J].眼外伤职业眼病杂志,2002,24(4):369-370.
[6]冷春生,李庆伟.牛胰蛋白酶制备工艺的改进及其对基因工程人胰岛素前体的酶切作用[J].沈阳药科大学学报,2013,30(1):68-71.
[7]张鹤,赵雨,徐云凤,等.胰蛋白酶水解梅花鹿鹿筋胶原工艺条件的研究[J].食品科技,2010,35(9):216-218.
[8]SAXENA S. Microbial enzymes and their industrial applications[M]//SAXENA S.Applied microbiology.India:Springer,2015:121-154.
[9]朱文辉,黄建强,张旭,等.胰蛋白酶水解蝇蛆蛋白粉的工艺研究[J].广东饲料,2014(7):27-29.
[10]国家药典委员会.中华人民共和国药典:二部[S].北京:化学工业出版社,2000:735-737.
[11]张锐,曾润颖.极端微生物产碱性蛋白酶菌株的筛选及发酵条件研究[J].微生物学通报,2001,28(4):5-9.
[12]CHANNE P S,SHEWALE J G.Influence of culture conditionson the formation of milk-clotting protease by Aspergillus niger MC4[J].World journal of microbiology and biotechnology,1998,14(1):11 15.
[13]孙倩,陈复生,丁长河,等.地衣芽孢杆菌产碱性蛋白酶发酵条件优化[J].食品工业科技,2012,33(13):177,192.