乳酸对地衣芽孢杆菌HK高密度发酵的影响  

Influences of Lactate on the High-density Fermentation by Bacillus licheniformis HK

作　　者：赵艳梅 鲁佳康 史玥玡 王彦婷 黄亚男 陈雄[1] 王志[1] ZHAO Yanmei;LU Jiakang;SHI Yueya;WANG Yanting;HUANG Yanan;CHEN Xiong;WANG Zhi(Key Laboratory of Fermentation Engineering(Ministry of Education),Collaborative Innovation Center for Industrial Fermentation,Hubei University of Technology,Wuhan 430068,China;Henan Province Nanjiecun Group Co.,Ltd.,Linying 462600,China)

机构地区：[1]发酵工程教育部重点实验室,工业发酵省部共建协同创新中心,湖北工业大学,湖北武汉430068 [2]河南省南街村(集团)有限公司,河南临颍462600

出　　处：《食品工业科技》2025年第7期151-160,共10页Science and Technology of Food Industry

基　　金：湖北省教育厅优秀中青年科技创新团队项目(T2022011)。

摘　　要：为了提高地衣芽孢杆菌HK(Bacillus licheniformis HK)的生物量和芽孢率,在20 L罐水平考察了发酵16~32 h耦合pH7.0流加乳酸(终浓度2.0%)对其生长代谢及产孢的影响,并基于转录组与代谢组差异分析了乳酸的代谢调控机制。结果表明:乳酸补料组的峰值生物量(30 h)为5.43×10^(10)CFU/mL,较对照(基础发酵组)提高54.3%;芽孢数为5.36×10^(10)CFU/mL,较对照提高60.0%。乳酸补料组的糖异生和磷酸戊糖途径关键基因ldh、pckA、gapB、zwf、tkt较对照上调2.8~9.3倍;同时,其标志代谢物丙酮酸、6-磷酸果糖、1,3-二磷酸甘油酸、6-磷酸葡萄糖酸、5-磷酸-核酮糖、7-磷酸景天庚酮糖的丰度较对照增加1.55~12.6倍。TCA循环和氧化磷酸化关键基因citZ、icd、citB、odhB、atpAB、ndh、yumB、qcrABC、ctaODC、ythA、sdhAB表达较对照下调34.2%~96.8%,而硝酸盐呼吸代谢基因nasBCD较对照上调107.4~287.3倍。与之相应的胞内关键代谢物顺乌头酸、异柠檬酸和柠檬酸丰度较对照降低52.6%~62.5%,而NAD+、NADH丰度较对照增加1.6和2.7倍。同时,磷壁酸的前体3-磷酸甘油、肽聚糖的前体UDP-N-乙酰胞壁酸和UDP-N-乙酰胞壁酰-L-丙氨酸-D-谷氨酸的丰度较对照提高了1.5~14倍;而且,芽孢生成相关基因phrAC、cotP、cotA、yeek、yheD、tasA、gerQ、yuzJ的表达较对照上调1.4~5.7倍,而rapAB、abrB、yisI、ynzD和spo0E较对照下调60.7%~96.7%。说明乳酸增强了糖异生、磷酸戊糖途径、硝酸盐呼吸、细胞壁合成,从而促进了细胞生长及芽孢的生成效率。本文结果为地衣芽孢杆菌高密度发酵生产提供了理论支撑。To improve the biomass and spore rate of Bacillus licheniformis HK,influences of lactate on cell growth and metabolism and spore production were investigated at a 20 L bioreactor,and lactate addition was conducted by coupling with pH7.0 from 16 h to 32 h.Basing on the data differences in transcriptome and metabolome,the metabolic regulation mechanism of lactate was analyzed.The results showed that a peak biomass(30 h)of 5.43×10^(10) CFU/mL was obtained in lactate supplement group,which was 54.3%higher than that of basic fermentation group(control).Meanwhile,the spore number was 5.36×10^(10) CFU/mL,which was 60.0%higher than the control.Compared with the control,expressions of key genes in gluconeogenesis and pentose phosphate pathway(ldh,pckA,gapB,zwf,and tkt)were up-regulated by 2.8~9.3 fold in the lactate supplement group,the abundance of marker metabolites(pyruvate,fructose-6-phosphate,glyceric acid-1,3-diphosphate,6-phosphate-gluconate,5-phosphate-ribulose,and sedoheptulose 7-phosphate)in the pathways were increased by 1.55~12.6 fold compared with the control.Additionally,expressions of key genes in TCA cycle and oxidative phosphorylation system(citZ,icd,citB,odhB,atpAB,ndh,yumB,qcrABC,ctaODC,ythA,and sdhAB)were down-regulated by 34.2%~96.8%,while nitrate respiration metabolism gene(nasBCD)was up-regulated by 107.4~287.3 fold compared with the control.Abundances of corresponding key metabolites,such as aconitate,isocitrate and citrate,were decreased by 52.6%~62.5%,while the abundances of NAD+and NADH were increased by 1.6~2.7 fold.Meanwhile,the precursors abundances of teichoic acid(glycerol-3-phosphate)and peptidoglycan(UDP-N-acetylmuramic acid,UDP-Nacetylmuramyl-L-alanine-D-glutamate)were increased by 1.5~14 fold compared with the control.Moreover,gene expressions involved in sporogenesis,such as phrAC,cotP,cotA,yeek,yheD,tasA,gerQ,yuzJ,were up-regulated by 1.4~5.7 fold,while rapAB,abrB,yisI,ynzD,and spo0E were down-regulated by 60.7%~96.7%compared with the control.The results suggested that lactate promo

关 键 词：地衣芽孢杆菌 乳酸 细胞生长 芽孢 转录组分析 代谢组分析 

分 类 号：TS201.3[轻工技术与工程—食品科学]