首页|外源添加VB12对肺炎克雷伯氏菌代谢3-羟基丙酸的影响

外源添加VB12对肺炎克雷伯氏菌代谢3-羟基丙酸的影响

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为了探究外源添加维生素 B12(Vitamin B12,VB12)对Klebsiella pneumoniae HD79 和K.pneumoniae HD79-T利用甘油还原途径生产3-羟基丙酸(3-hydroxypropionic acid,3-HP)的影响以及摸索VB12对K.pneumoniae HD79和K.pneumoniae HD79-T的阈值上限,将不同浓度(0.01、0.02、0.03、0.04、0.05 g/L)的VB12添加到K.pneumoniae HD79及K.pneumoniae HD79-T的发酵培养基中,利用HPLC检测其底物消耗及产物产生情况、qRT-PCR检测还原途径相关基因的mRNA表达情况以及酶联免疫试剂盒检测代谢相关酶活性.结果表明,VB12对K.pneumoniae HD79和K.pneumoniae HD79-T的阈值为0.01 g/L和0.03 g/L.与未添加VB12相比,菌株K.pneumoniaeHD79和K.pneumoniaeHD79-T的3-HP产量分别提高了 24.39％,8.86％;醛脱氢酶基因puuC表达量分别提高了2.49倍和1.68倍;ALDH、GDHt和PDOR的酶活力分别提高了 50.24％、40.36％和18.29％,及30.49％、37.84％和13.56％.说明通过外源添加辅酶因子VB12对肺炎克雷伯氏菌高产3-HP是可行策略.
Effect on Metabolism of 3-hydroxypropionic Acid by Klebsiella pneumoniae:Exogenous Addition of VB12
To investigate the effect of exogenous addition of Vitamin B12(VB112)on the production of 3-hydroxypropionic acid(3-HP)through glycerol reduction pathway in Klebsiella pneumoniae HD79 and K.pneumoniae HD79-T,and to explore the threshold upper limit of VB12 on the above strains,different concentrations(0.01,0.02,0.03,0.04,and 0.05 g/L)of VB12 were added to the fermentation medium of K.pneumoniae HD79 and K.pneumoniae HD79-T.The consumption of substrates and production of products were detected by HPLC,mRNA expression of genes related to the reduction pathway was detected by qRT-PCR,and enzyme activities related to metabolism were detected by enzyme-linked immunosorbent assay(ELISA).The results showed that the thresholds of VB12 for K pneumoniae HD79 and K.pneumoniae HD79-T was 0.01 g/L and 0.03 g/L,respectively.Compared with the absence of VB12,the productions of 3-HP of K pneumoniae HD79 and K.pneumoniae HD79-T were increased by 24.39％and 8.86％,respectively;the expression levels of aldehyde dehydrogenase gene puuC were increased by 2.49-fold and 1.68-fold,respectively;the enzyme activities of ALDH,GDHt,and PDOR increased by 50.24％,40.36％,and 18.29％in K pneumoniaeHD79,and by 30.49％,37.84％,and 13.56％in K.pneumoniae HD79-T,respectively.These results indicated that exogenous addition of coenzyme factor VB12 was a feasible strategy for high production of 3-HP in Klebsiella pneumoniae.

Vitamin B12Klebsiella pneumoniae3-hydroxypropionic acidaldehyde dehydrogenase gene puuCglycerol dehydratase

敖国旭、田甜、葛菁萍、平文祥

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黑龙江大学,生命科学学院,农业微生物技术教育部工程研究中心,黑龙江省寒区植物基因与生物发酵重点实验室,黑龙江省普通高校分子生物学重点实验室,哈尔滨 150080

河北环境工程学院,河北省农业生态安全重点实验室,河北秦皇岛 066102

维生素B12 肺炎克雷伯氏菌 3-羟基丙酸 醛脱氢酶基因puuC 甘油脱水酶

黑龙江省自然科学基金重点项目黑龙江省农业废弃物综合利用体系岗位专家项目黑龙江省生态环境厅2022年度生态环境保护科研项目龙江省高校基本科研业务费

ZD2020C008HST2022TR0042023-KYYWF-1448

2024

中国农学通报
中国农学会

中国农学通报

CSTPCD
影响因子：0.891
ISSN：1000-6850
年,卷(期)：2024.40(12)
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