食品科学

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产蛋白酶兼性厌氧菌株的筛选、酶学性质及发酵豆粕应用探究

熊 涛,唐晓星,黄 涛,李 啸   

  1. 南昌大学生命科学与食品工程学院,食品科学与技术国家重点实验室,江西 南昌 330047
  • 出版日期:2014-05-15 发布日期:2014-05-19

Screening of Protease-Producing Bacterium in Facultative Anaerobic Conditions, Studies on Its Enzymatic Characterization and Application for Fermenting Soybean Meal

XIONG Tao, TANG Xiao-xing, HUANG Tao, LI Xiao   

  1. State Key Laboratory of Food Science and Technology, School of Life Sciences and Food Engineering,Nanchang University, Nanchang 330047, China
  • Online:2014-05-15 Published:2014-05-19

摘要:

为筛选得到1 株或几株深层发酵豆粕效果较好的菌株,以透明圈法从自然发酵的豆豉中筛选出4 株兼性厌氧且产蛋白酶菌株,同时发现菌株B-1、B-12在厌氧条件下产蛋白酶能力好于好氧条件,对此现象较明显且平板水解圈直径较大的菌株B-1进行形态学、生理生化和分子生物学实验,鉴定为甲基营养型芽孢杆菌。进一步对菌株B-1的蛋白酶酶学性质进行研究后发现,酶活力在pH 6.0~9.0的范围内能够维持一个较高的水平,酶反应和酶稳定最适条件为pH 7.0和40 ℃,加入Mn2+对酶反应的促进作用较大。将甲基营养型芽孢杆菌B-1以6%的接种量接种于豆粕固态发酵培养基中堆积(20 cm)发酵,以枯草芽孢杆菌A-12好氧浅层发酵和未发酵豆粕作为对照,发酵结束后测得豆粕中小肽含量和水解度分别高达28.37%和29.55%,比枯草芽孢杆菌固态发酵对照组分别高出了9.04%和16.07%,与未发酵豆粕相比,菌株B-1发酵后豆粕的小肽含量和水解度也分别提高了27.22%和29.13%。

关键词: 蛋白酶, 兼性厌氧, 甲基营养型芽孢杆菌, 鉴定, 酶学性质, 发酵豆粕

Abstract:

An efficient protease–producing and facultative anaerobic bacterium B-1 was isolated from naturally fermented
Douchi by transparent inhibition zone plate assay. The diameter of inhibition zones in anaerobic conditions was found to be
bigger than in aerobic conditions. Based on physiological and biochemical identification and 16S rRNA sequence analysis, it
was identified as Bacillus methylotrophicus. Meanwhile, this study revealed that the optimal pH and temperature for protease
activity and stability from Bacillus methylotrophicus B-1 were pH 7.0 and 40 ℃, respectively. The protease activity was
activated by Mn2+. In addition, Bacillus methylotrophicus B-1 and Bacillus subtilis A-12 were used at an inoculum size of 6%
on raw soybean meal to study the effect of fermentation on the contents of crude protein and small peptides and hydrolysis
degree. The results showed that the content of small peptides and hydrolysis degree of soybean meal fermented by Bacillus
methylotrophicus B-1 were 28.37% and 29.55%, which were 9.04% and 16.07% higher than those of that fermented by
Bacillus subtilis A-12, and 27.22% and 29.13% higher than those of raw soybean meal, respectively.

Key words: protease, facultative anaerobic, Bacillus methylotrophicus, identification, enzymatic properties, fermented soybean meal