脉冲磁场诱变结合发酵条件优化提高中性蛋白酶产量的研究
|
摘要
利用脉冲磁场诱变枯草芽孢杆菌结合发酵条件优化以实现中性蛋白酶产量的最大化。结果表明在磁场强度为7 T,脉冲数为40次时诱变得到了中性蛋白酶产量提高17.7%的枯草芽孢杆菌菌株。对其发酵条件进行优化,得到最佳发酵培养基配方为豆粕粉90 g/L,麦芽糖10 g/L,KH_2PO_4 8 g/L,最佳发酵条件为接种量5%、发酵温度37℃、摇床转速180 r/min。在此条件下,发酵48 h,中性蛋白酶酶活力为384.57 U/m L,比野生株提高了26.48%。
The pulsed magnetic field was used to induce mutagenesis of Bacillus subtilis in combination with optimization of fermentation conditions to maximize the production of neutral protease. Results show that when the magnetic field intensity was 7 T and the number of pulses was 40,the Bacillus subtilis strains were obtained by mutagenesis with an increase of 17.7% in neutral protease production.After the fermentation conditions were optimized,the optimum fermentation medium formulation was obtained as follows:soybean meal 90 g/L,maltose 10 g/L,KH_2PO_4 8 g/L,and the optimum fermentation conditions were inoculation 5%,fermentation temperature 37 ℃,and shaker speed 180 r/min. After fermentation of 48 h under these conditions,the activity of the neutral protease was 384.57 U/mL,an increase of 26.48 % against the wild strains.
The pulsed magnetic field was used to induce mutagenesis of Bacillus subtilis in combination with optimization of fermentation conditions to maximize the production of neutral protease. Results show that when the magnetic field intensity was 7 T and the number of pulses was 40,the Bacillus subtilis strains were obtained by mutagenesis with an increase of 17.7% in neutral protease production.After the fermentation conditions were optimized,the optimum fermentation medium formulation was obtained as follows:soybean meal 90 g/L,maltose 10 g/L,KH_2PO_4 8 g/L,and the optimum fermentation conditions were inoculation 5%,fermentation temperature 37 ℃,and shaker speed 180 r/min. After fermentation of 48 h under these conditions,the activity of the neutral protease was 384.57 U/mL,an increase of 26.48 % against the wild strains.
引文
[1]张晓燕,国立东,刘晓艳.枯草芽孢杆菌中性蛋白酶的研究进展[J].中国酿造,2018,37(4):12-15.
[2]Lu Zhen,Jia Chenfu,Li Enzhong,et al.Screening and Isolation of High Neutral Protease Yielding Strains and Study on the Best Fermentation Performance[J].Heilongjiang Agricultural Sciences,2016(5):110-113.
[3]高产谷氨酸北京棒杆菌诱变育种[D].黑龙江大学,2013.
[4]曲文娟,张天,马海乐,等.高产蛋白酶菌株等离子体诱变及其在提高豆粕发酵效果中的应用[J].现代食品科技,2018,34(10):133-140.
[5]张雪娇,李淑娥,谭利,等.花生四烯酸高产菌株的脉冲强磁场诱变选育[C]//武汉:湖北省暨武汉微生物学会会员代表大会暨学术年会,2013.
[6]黄达明,宋庆春,管国强,等.枯草芽孢杆菌ZC1发酵产中性蛋白酶的条件优化[J].中国调味品,2015(9):35-40.
[7]刘颖,张彬彬,孙冰玉,等.枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J].食品科学,2014,35(13):166-170.
[8]张士伟,黄建飞,罗立新.枯草芽孢杆菌产蛋白酶发酵培养基的优化[J],中国酿造,2013,32(2):20-24.
[9]刘新风,牛春华,刘香英,等.枯草芽孢杆菌BSG1产蛋白酶发酵条件优化[J].食品工业,2013(7):1-4.
[10]刘晓艳,张晓燕,杨国力,等.枯草芽孢杆菌L1发酵产中性蛋白酶活性的研究[J].食品与发酵科技,2018,54(3):55-59.
[11]许曌昕,曾辉,曾德勇,等.响应面法优化解淀粉芽孢杆菌产中性蛋白酶发酵条件[J].中国酿造,2017,36(4):78-82.
[12]刘丽娜,王安建,魏书信,等.乳酸菌发酵香菇柄酶解物产γ-氨基丁酸的研究[J].包装与食品机械,2018,36(4):13-19.
[13]王东,荣家萍,唐自钟,等.响应面法优化枯草芽孢杆菌产中性蛋白酶的发酵条件[J].基因组学与应用生物学,2016,35(1):143-151.
[14]刘颖,张彬彬,孙冰玉,等.枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J].食品科学,2014,35(13):166-170.
[15]王东,荣家萍,唐自钟,等.响应面法优化枯草芽孢杆菌产中性蛋白酶的发酵条件[J].基因组学与应用生物学,2016,35(1):143-151.
[16]李洪康,李由然,李赢,等.枯草芽孢杆菌产中性蛋白酶发酵条件优化[J].食品与发酵工业,2016,42(5):102-107.
[2]Lu Zhen,Jia Chenfu,Li Enzhong,et al.Screening and Isolation of High Neutral Protease Yielding Strains and Study on the Best Fermentation Performance[J].Heilongjiang Agricultural Sciences,2016(5):110-113.
[3]高产谷氨酸北京棒杆菌诱变育种[D].黑龙江大学,2013.
[4]曲文娟,张天,马海乐,等.高产蛋白酶菌株等离子体诱变及其在提高豆粕发酵效果中的应用[J].现代食品科技,2018,34(10):133-140.
[5]张雪娇,李淑娥,谭利,等.花生四烯酸高产菌株的脉冲强磁场诱变选育[C]//武汉:湖北省暨武汉微生物学会会员代表大会暨学术年会,2013.
[6]黄达明,宋庆春,管国强,等.枯草芽孢杆菌ZC1发酵产中性蛋白酶的条件优化[J].中国调味品,2015(9):35-40.
[7]刘颖,张彬彬,孙冰玉,等.枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J].食品科学,2014,35(13):166-170.
[8]张士伟,黄建飞,罗立新.枯草芽孢杆菌产蛋白酶发酵培养基的优化[J],中国酿造,2013,32(2):20-24.
[9]刘新风,牛春华,刘香英,等.枯草芽孢杆菌BSG1产蛋白酶发酵条件优化[J].食品工业,2013(7):1-4.
[10]刘晓艳,张晓燕,杨国力,等.枯草芽孢杆菌L1发酵产中性蛋白酶活性的研究[J].食品与发酵科技,2018,54(3):55-59.
[11]许曌昕,曾辉,曾德勇,等.响应面法优化解淀粉芽孢杆菌产中性蛋白酶发酵条件[J].中国酿造,2017,36(4):78-82.
[12]刘丽娜,王安建,魏书信,等.乳酸菌发酵香菇柄酶解物产γ-氨基丁酸的研究[J].包装与食品机械,2018,36(4):13-19.
[13]王东,荣家萍,唐自钟,等.响应面法优化枯草芽孢杆菌产中性蛋白酶的发酵条件[J].基因组学与应用生物学,2016,35(1):143-151.
[14]刘颖,张彬彬,孙冰玉,等.枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J].食品科学,2014,35(13):166-170.
[15]王东,荣家萍,唐自钟,等.响应面法优化枯草芽孢杆菌产中性蛋白酶的发酵条件[J].基因组学与应用生物学,2016,35(1):143-151.
[16]李洪康,李由然,李赢,等.枯草芽孢杆菌产中性蛋白酶发酵条件优化[J].食品与发酵工业,2016,42(5):102-107.