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谷氨酸棒杆菌中L-高丝氨酸转运蛋白的挖掘及功能分析
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作者 张霞 祁玉婷 +4 位作者 钟兆月 徐宁 刘君 姜巨全 邵丽 《食品与发酵工业》 北大核心 2025年第4期34-41,共8页
L-高丝氨酸是一种非必需手性氨基酸,在农业、化工、生物医药等领域具有重要的用途。近年来,利用微生物发酵法生产L-高丝氨酸越来越受到人们的关注。氨基酸转运蛋白不仅可以降低反馈抑制和细胞毒性,还能提高氨基酸产量,已成为代谢工程改... L-高丝氨酸是一种非必需手性氨基酸,在农业、化工、生物医药等领域具有重要的用途。近年来,利用微生物发酵法生产L-高丝氨酸越来越受到人们的关注。氨基酸转运蛋白不仅可以降低反馈抑制和细胞毒性,还能提高氨基酸产量,已成为代谢工程改造的重要靶点。目前,谷氨酸棒杆菌中L-高丝氨酸转运蛋白仍然未知。该研究通过生物信息学分析,从谷氨酸棒杆菌中筛选出10个L-高丝氨酸转运蛋白候选基因,经过体外和体内L-高丝氨酸耐受性生长实验,结果显示在大肠杆菌三重高丝氨酸转运缺陷突变体中,异源过表达brnFE、azlCD和lysE基因,能显著恢复大肠杆菌突变体的生长缺陷,而在谷氨酸棒杆菌体内,只有lysE敲除菌较对照生物量下降16%,推测在体内LysE蛋白具有更重要的高丝氨酸外排能力。进一步对LysE的外排活性进行分析,发现lysE敲除菌的外排能力降低了18%。最后,在高丝氨酸底盘菌中,评估了敲除或过表达lysE基因对L-高丝氨酸产量的影响,结果发现lysE基因缺失导致L-高丝氨酸的产量降低了65%,而过表达lysE使L-高丝氨酸产量提高了22%。综上所述,谷氨酸棒杆菌LysE蛋白具有转运L-高丝氨酸的能力,相关结果为开发高效的L-高丝氨酸细胞工厂提供了新靶点。 展开更多
关键词 L-高丝氨酸 外排蛋白 谷氨酸棒杆菌 序列比对 LysE
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Transformation of organic matters in fresh leachate during anaerobic degradation under long hydraulic retention time 被引量:5
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作者 XU Suyun,HE Pinjing,LU Fan,LI Min,SHAO Liming State Key Laboratory of Pollution Control and Resources Reuse,Key Laboratory of Yangtze River Water Environment,College of Environmental Science and Engineering,Tongji University,Shanghai 200092,China 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2009年第9期1302-1308,共7页
This study investigated the biodegradability of fresh leachate from a young municipal solid waste landfill at the inoculum to substrate ratios (V/V) of 10/100, 25/100, 50/100 and 100/100, as well as the transformati... This study investigated the biodegradability of fresh leachate from a young municipal solid waste landfill at the inoculum to substrate ratios (V/V) of 10/100, 25/100, 50/100 and 100/100, as well as the transformation of organic matters in leachate under prolonged hydraulic retention time (HRT) of 50 d. Fresh leachate showed a good biodegradability, with chemical oxygen demand (COD) removal efficiency as high as 87%-92% and cumulative methane yield close to theoretical value (0.35 L CH4/g CODdeg). Methane production and COD depletion presented biphasic characteristics associated with the successive utilization of two major intermediates, acetate and propionate, The biphasic degradation of fresh leachate was resulted from the different hydrolysis rates of diverse substrates and the changes in microorganism community structure. After 50 d, the effluents were dominated by high-MW organic compounds (MW 〉 10 kDa) at each inoculum ratio, which might be refractory compounds released from cell lyses. 展开更多
关键词 BIODEGRADABILITY biomass lyses methane production ratio of inoculum to substrate volatile fatty acids
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The algae-lytic ability of bacterium DC_(10) and the influence of environmental factors on the ability
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作者 SHI Shunyu 1,2 , LIU Yongding 1 , SHEN Yinwu 1 & LI Genbao 1 1. Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China 2. Graduate School of Chinese Academy of Sciences, Beijing 100039, China 《Science China(Life Sciences)》 SCIE CAS 2005年第3期250-255,共6页
A lysing-bacterium DC10, isolated from Dianchi Lake of Yunnan Province, was characterized to be Pseudomonas sp. It was able to lyse some algae well, such as Microcystis viridis, Selenastrum capricornutum, and so on. I... A lysing-bacterium DC10, isolated from Dianchi Lake of Yunnan Province, was characterized to be Pseudomonas sp. It was able to lyse some algae well, such as Microcystis viridis, Selenastrum capricornutum, and so on. In this study, it was shown that the bacterium lysed the algae by releasing a substance; the best lytic effects were achieved at low tempera- tures and in the dark. Different concentrations of CaCl2 and NaNO3 influenced the lytic effects; the ability to lyse algae decreased in the following order: pH 4 > pH 9 > pH 7 > pH 5.5. It was significant to develop a special technology with this kind of bacterium for controlling the bloom- forming planktonic microalgae. 展开更多
关键词 lysing-bacterium lyse algae control.
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毛萼田菁茎瘤菌被种子浸提物诱导表达基因的筛选及其功能
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作者 蔡文通 蔡韬 +3 位作者 张江 郑会明 钟增涛 朱军 《微生物学报》 CAS CSCD 北大核心 2009年第9期1171-1175,共5页
【目的】选择毛萼田菁茎瘤菌ORS571为研究对象,筛选毛萼田菁茎瘤菌在菌植互作早期被宿主植物种子浸提物诱导表达的基因。【方法】采用新颖的基于抗性的活体表达筛选技术,以种子浸提物为诱导物,筛选获得能够稳定被种子浸提物诱导的突变... 【目的】选择毛萼田菁茎瘤菌ORS571为研究对象,筛选毛萼田菁茎瘤菌在菌植互作早期被宿主植物种子浸提物诱导表达的基因。【方法】采用新颖的基于抗性的活体表达筛选技术,以种子浸提物为诱导物,筛选获得能够稳定被种子浸提物诱导的突变株。通过中间片断融合报告基因的方法研究信号分子对相关基因的诱导情况。【结果】随机引物PCR产物测序及网上BLAST结果表明,被诱导表达的基因为LysE家族的成员。而且,对另外3个属根瘤菌中lysE基因的研究发现,它们都可以被相应的宿主植物种子浸提物诱导表达。此外,这四株根瘤菌的lysE突变株都表现对刀豆氨酸极为敏感。本文首次报道了lysE家族在根瘤菌中的生理功能。【结论】我们推测:LysE家族蛋白可能广泛存在于根瘤菌中,并在根瘤菌与宿主植物的早期相互作用中发挥重要作用。 展开更多
关键词 毛萼田菁茎瘤菌 LysE家族 刀豆氨酸 种子浸提物
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