耐盐植物乳杆菌的选育及其对高盐稀态发酵酱油品质的影响  

Breeding of salt-tolerant Lactobacillus plantarum and their effects on the quality of high-salt liquid-state fermented soy sauce

作　　者：张伟[1] 杨俊文 余冰艳 林琛 谢梦雪 罗晓明 周尚庭 蒋雪薇[1,3] ZHANG Wei;YANG Junwen;YU Bingyan;LIN Chen;XIE Mengxue;LUO Xiaoming;ZHOU Shangting;JIANG Xuewei(School of Food Science and Bioengineering,Changsha University of Science and Technology,Changsha 410114,China;School of Chemistry and Chemical Engineering,Changsha University of Science and Technology,Changsha 410114,China;Hunan Provincial Engineering Technology Research Center for Condiment Fermentation,Changsha 410600,China;Jiajia Food Group Co.Ltd.,Changsha 410600,China)

机构地区：[1]长沙理工大学食品与生物工程学院,湖南长沙410114 [2]长沙理工大学化学化工学院,湖南长沙410114 [3]湖南省调味品发酵工程技术研究中心,湖南长沙410600 [4]加加食品集团股份有限公司,湖南长沙410600

出　　处：《食品与发酵工业》2023年第1期86-94,共9页Food and Fermentation Industries

基　　金：湖南省自然科学基金面上项目(2021 JJ30700);湖南省教育厅重点项目(21A0197);长沙市科技计划重点研发项目(kh200595,kq2004072);长沙理工大学研究生实践创新与创业能力提升计划项目(SJCX202187)。

摘　　要：为得到发酵性能优良且适用于高盐稀态酱油酿造的耐盐乳酸菌,以发酵泡菜中筛选的耐盐植物乳杆菌(Lactobacillus plantarum)L04为出发菌株进行紫外诱变,高锰酸钾-溴化钾-MRS(de Man,Rogosa and Sharpe)平板初筛及MRS液体发酵复筛获得2株产乳酸较强的突变株L04-1和L04-2,其在160 g/L NaCl的MRS液体培养基中发酵乳酸积累量分别可达3.69和3.49 g/L,比L04高16.03%和9.75%。在高盐稀态发酵第45天添加L04-1及L04-2进行为期120 d的发酵,乳酸菌总数和细菌总数的测定结果表明,耐盐植物乳杆菌的加入可增加酱醪中乳酸菌的数量并能抑制其他细菌生长;通过对酱油发酵过程中乳酸、还原糖、总酸及氨态氮的分析发现,2株突变株均可促进乳酸、总酸及氨态氮的积累,其中添加L04-1发酵酱油乳酸含量最高,为4.31 g/L。采用气质联用分析添加L04-1、L04-2发酵酱油的挥发性风味物质,发现其酯类物质含量分别比L04高36.60%和16.46%;醇类物质分别比L04高23.24%和10.58%,且含量均高于对照组,可促进酱油的前香及主体香的形成。感官评分显示,2株突变株发酵的酱油香气和滋味的评分均优于添加L04的发酵酱油和对照组。诱变选育的2株耐盐植物乳杆菌在酱油发酵中具有积累乳酸、促进酯类、醇类物质生成的特征,有利于形成酱油适口的酸感,丰富酱油的滋味和香气。In order to obtain salt-tolerant lactic acid bacteria with excellent fermentation performance and suitable for high-salt liquid-state soy sauce brewing,salt-tolerant Lactobacillus plantarum L04,isolated from fermented kimchi,was treated by ultraviolet mutagenesis.And two mutants(L04-1,L04-2)with stronger lactic acid production were obtained by primary screening in Man Rogosa Sharp(MRS)plates with KMnO 4 and KBr,and secondary screening with MRS liquid fermentation.They could accumulate 3.69 and 3.49 g/L lactic acid in MRS liquid medium with 160 g/L NaCl,respectively,which were 16.03%and 9.75%higher than L04.On the 45 th day of high-salt liquid-state fermentation,L04-1 and L04-2 were added in moromi for 120 d of fermentation.Results showed that the addition of salt-tolerant L.plantarum could increase the lactic acid bacteria counts while inhibit the growth of other bacteria in the moromi.Two mutants could promote the accumulation of lactic acid,total acid and ammonia nitrogen.The highest lactic acid content in soy sauce fermented with L04-1 was 4.31 g/L.The volatile flavor compounds of soy sauce fermented with L04-1 and L04-2 were analyzed by GC-MS.The ester content of soy sauce fermented with two mutants was 36.60%and 16.46%higher than the original L04 strain,and the alcohol content was 23.24%and 10.58%higher than L04,respectively.The ester and alcohol content were also higher than the control,which indicated that the two mutants can promote the formation of top and main aromas of soy sauce.Aroma and taste scores of soy sauce fermented by the two mutants were better than soy sauce fermented with L04 and the control.The two strains of salt-tolerant L.plantarum bred by ultraviolet mutagenesis could accumulate lactic acid and promote the production of esters and alcohols in soy sauce fermentation,which can be applied in soy sauce brewing to enhance the palatable acidity and enrich its taste and aroma.

关 键 词：耐盐植物乳杆菌 选育 高盐稀态发酵酱油 乳酸 挥发性风味物质 

分 类 号：TS264.21[轻工技术与工程—发酵工程]