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High Technology Letters 2004年第4期10卷 69-74页

作者：王相晶 Ren Nanqi xiang wensheng Life Science College Northeast Agricultural University 150030 Harbin P.R.China College of Municipal & Environmental Engineering Harbin Institute of Technology 150090 Harbin P.R.China

Iron plays an important role in hydrogen production, cell growth, hydrogenase and NADH-fd reductase activities of hydrogen-producing bacterial strain B49 (AF481148 in EMBL). At the end of fermentation from 10 g/L glucose, for the culture containing 10 mg/L FeSO4*7H2O the cell growth in terms of optical density (OD) at 600nm was 1.13, the ratio of ethanol amount (mg/L) to acetate amount (mg/L) was 1.55, and the accumulated hydrogen volume was 1816.3 ml H2/L culture; whereas for the culture of 80 mg/L FeSO4*7H2O OD600nm was increased to 1.34, the accumulated hydrogen volume was increased to 2360.5 ml H2/L culture, and the ratio of ethanol amount (mg/L) to acetate amount (mg/L) decreased to 1.31. Moreover, the iron addition to the medium at different fermentation time could affect hydrogen-producing ability. However, the later the addition time of FeSO4*7H2O was postponed, the less the effect on hydrogen evolution was. In the course of fermentation, the specific activities of hydrogenase and NADH-fd reductase of hydrogen-producing bacterial strain B49 decreased with the consumption of iron.

关键词：氢化酶还原酶生物活性发酵工艺细菌 NADH-fd 能源清洁燃料生物燃料

The start-up of biohydrogen-producing process by bioaugmentation in the EGSB reactor

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High Technology Letters 2006年第3期12卷 328-332页

作者：王相晶 Ren Nanqi xiang wensheng Guo Wanqian School of Municipal ＆ Environmental Engineering Harbin Institute of Technology Harbin 150090 P.R. China School of Life Science Northeast Agricultural University Harbin 150030 P.R. China

Expanded granular sludge bed （EGSB） reactor and bioaugmentation were employed to investigate biohydrogen production with molasses wastewater. The start-up experiments consisted of two stages. In the first stage （0 - 24d） seeded with activated sludge, the butyric acid type-fermentation formed when the initial expanding rate, organic loading rate （OLR）, the initial redox potential （ORP） and hydraulic retention time （HRT） were 10%, 10.0 kg COD/（m^3·d）, -215 mV and 6.7 h, respectively. At the beginning of the second stage on day 25, the novel hydrogen-producing fermentative bacterial strain B49 （AF481148 in EMBL） were inoculated into the reactor under the condition of OLR 16. 0 kg COD/（m^3·d）, ORP and HRT about - 139 mV and 6.7 h, respectively, and then the reaction system transformed to ethanol-type fermentation gradually with the increase in OLR. When OLR, ORP and HRT were about 94.3 kg COD/（m^3·d）, -250 mV and 1.7 h, respectively, the system achieved the maximum hydrogen-producing rate of 282.6 mL H2/L reactor·h and hydrogen percentage of 51% -53% in the biogas.

关键词： start-up biohydrogen production bioaugmentation hydrogen-producing bacterial strain B49 EGSB reactor

Mining and fine-tuning sugar uptake system for titer improvement of milbemycins in Streptomyces bingchenggensis

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Synthetic and Systems Biotechnology 2020年第3期5卷 214-221页

作者： Pinjiao Jin Shanshan Li Yanyan Zhang Liyang Chu Hairong He Zhuoxu Dong wensheng xiang School of Life Science Northeast Agricultural UniversityNo.59 Mucai StreetXiangfang DistrictHarbin150030China State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing100193China College of Pharmacy Henan University of Chinese MedicineZhengzhou 450046HenanChina

Dramatic decrease of sugar uptake is a general phenomenon in Streptomyces at stationary phase,when antibiotics are extensively *** produced by Streptomyces bingchenggensis are a group of valuable macrolide biopesticides,while the low yield and titer impede their broad applications in agricultural *** that inadequate sugar uptake generally hinders titer improvement of desired products,we mined the underlying sugar uptake systems and fine-tuned their expression in this ***,we screened the candidates at both genomic and transcriptomic level in ***,two ATP-binding cassette transporters named TP2 and TP5 were characterized to improve milbemycin titer and yield ***,the appropriate native temporal promoters were selected and used to tune the expression of TP2 and TP5,resulting in a maximal milbemycin A3/A4 titer increase by 36.9%to 3321 mg/***,TP2 and TP5 were broadly finetuned in another two macrolide biopesticide producers Streptomyces avermitilis and Streptomyces cyaneogriseus,leading to a maximal titer improvement of 34.1%and 52.6%for avermectin B1a and nemadectin,*** work provides useful transporter tools and corresponding engineering strategy for Streptomyces.

关键词： Streptomyces Sugar uptake system Fine-tuning Titer improvement Milbemycins Macrolide biopesticides

Hydrogen Production with High Evolution Rate and High Yield by Immobilized Cells of Hydrogen-producing Bacteria Strain B49 in a Column Reactor

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High Technology Letters 2002年第4期8卷 21-25页

作者：任南琪 Wang xiangjing xiang wensheng Lin Ming Li Jianzheng Guo Wanqian School of Municipal & Environmental Engineering Harbin Institute of Technology Harbin 150090 P.R China Research Center of Life Science and Biotechnology Northeast Agricultural University Harbin 150030 P.R China

To improve the hydrogen evolution rate in continuous hydrogen production of a fermentative hydrogen-producing bacteria strain B49 (AF481148 in EMBL), 4% immobilized cells by polyvinyl alcohol-boric acid method, with the addition of a small amount of calcium alginate in a column reactor obtain hydrogen yield of 2.31 mol H2/mol glucose and hydrogen evolution rate of 1435.4 ml/L culture·h respectively at medium retention time of 2 h with a medium containing l0 g glucose/L. As the cell density in gel beads is increased to 8%, hydrogen yield and hydrogen evolution rate for l0 g glucose/L are 2.34 mol H2/mol glucose and 2912.4 ml/L culture · h respectively at medium retention time of 1 h, and for molasses wastewater COD of 7505.9 mg/L hydrogen production potential of 205.6 ml/g COD and hydrogen evolution rate of 2057.7 ml/L culture·h at hydraulic retention time of 0.75 h are observed. In the continuous culture pH value keeps around 3.9 by self regulation.

关键词： hydrogen-producing bacteria strain B49,immobilized cells,continuous hydrogen production

Transcriptome-guided identification of a four-component system, SbrH1-R, that modulates milbemycin biosynthesis by influencing gene cluster expression, precursor supply, and antibiotic efflux

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Synthetic and Systems Biotechnology 2022年第2期7卷 705-717页

作者： Lan Ye Yanyan Zhang Shanshan Li Hairong He Guomin Ai xiangjing Wang wensheng xiang School of Life Science Northeast Agricultural UniversityNo.59 Mucai StreetXiangfang DistrictHarbin150030China State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing100193China State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijingChina College of Pharmacy Henan University of Chinese MedicineZhengzhou 450046HenanChina

Streptomyces can produce numerous antibiotics and many other bioactive ***,the molecular mechanisms of transcriptional regulators in control of antibiotic production by influencing the expression of biosynthetic gene clusters(BGCs)have been extensively ***,for regulators that affect both antibiotic production and cell growth,the way to influence antibiotic production may be diverse,but related studies are ***,based on time-course transcriptome analysis,a four-component system,SbrH1-R,consisting of the two-component system SbrKR(SBI_03479/3478)and two hypothetical proteins SbrH1(SBI_03481)and SbrH2(SBI_03480)potentially related with the biosynthesis of milbemycins was identified in Streptomyces bingchenggensis *** of sbrH1-R resulted in weakened cell growth but a 110%increase of milbemycin production compared with that in *** transcriptome analyses of the sbrH1-R mutant and BC-101-4 revealed that SbrH1-R not only indirectly represses milbemycin BGC expression,but also inhibits milbemycin production by modulating expression levels of genes related to precursor supply and antibiotic *** genetic experiments identified several new targets,including five precursor supply-associated reactions/pathways(e.g.,the reaction from pyruvate to acetyl-CoA,the reaction from acetyl-CoA to citrate,the fatty acidβ-oxidation process,and the branched chain amino acid and phenylalanine acid degradation pathways)and a milbemycin exporter system(MilEX2)that can be engineered for milbemycin *** results shed new light on the understanding of regulation of milbemycin biosynthesis and provide useful targets for future metabolic engineering of the native host to improve milbemycin production.

关键词： Milbemycins Streptomyces bingchenggensis SbrH1-R Precursor supply Milbemycin exporter Overproduction

Characterization of a LAL-type regulator NemR in nemadectin biosynthesis and its application for increasing nemadectin production in Streptomyces cyaneogriseus

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Science China(Life Sciences) 2019年第3期62卷 394-405页

作者： Chuang Li Hairong He Jiabin Wang Hui Liu Haiyan Wang Yajie Zhu xiangjing Wang Yanyan Zhang wensheng xiang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences School of Life Science Northeast Agricultural University College of Food and Bioengineering Qiqihar University

Nemadectin, a macrocyclic lactone antibiotic, is produced by Streptomyces cyaneogriseus ssp. noncyanogenus. A methoxime derivative of nemadectin, moxdectin, has been widely used to control insect and helminth in animal health. Despite the importance of nemadectin, little attention has been paid to the regulation of nemadectin biosynthesis, which has hindered efforts to improve nemadectin production via genetic manipulation of regulatory genes. Here, we characterize the function of nemR, the cluster-situated regulatory gene encoding a LAL-family transcriptional regulator, in the nemadectin biosynthesis gene cluster of S. cyaneogriseus ssp. noncyanogenus NMWT1. NemR is shown to be essential for nemadectin production and found to directly activate the transcription of nemA1-1/A1-2/A2, nemC and nemA4/A3/E/D operons, but indirectly activate that of nemG and nemF. A highly conserved sequence 5′-TGGGGTGKATAGGGGGTA-3′(K=T/G) is verified to be essential for NemR ***, four novel targets of NemR, including genes encoding an SsgA-like protein(TU94_12730), a methylmalonyl-CoA mutase(TU94_19950), a thioesterase of oligomycin biosynthesis(TU94_22425) and a MFS family transporter(TU94_24835)are identified. Overexpression of nemR significantly increased nemadectin production by 79.9%, in comparison with NMWT1,suggesting that nemR plays an important role in the nemadectin biosynthesis.

关键词： nemadectin Streptomyces cyaneogriseus ssp.noncyanogenus nemR LAL overexpression

Trace Residue Analysis of the Herbicide Imazethapyr in Soil by Gas Chromatography-Nitrogen/Phosphorous Detector

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Journal of Northeast Agricultural University(English Edition) 1997年第1期4卷 73-77页

作者： xiang wensheng, Su Shaoquan, Wu Yongqing and Han Xijiang (Northeast Agricultural University, Harbin 150030, PRC)

A gas chromatographic method for determining the trace residue of imazethapyr {(±)-2-[4,5-dihydro-4-methtyl-4-(1-methylethyl)-5-oxo-1H-imidazol-2-yl]-5-ehtyl-3-pyridinecarboxylic acid} in soil was described in this paper. The soil samples were extracted by shaking with 0.5 mol·L -1 sodium hydroxide solution. The aqueous extract was acidified with hydrocholric acid to adjust the pH to 2.0 to precipitate humic acids, then was extracted with dichloromethane. The organic extract was dried with anhydrous sodium sulfate and was concentrated. After florisil cleanup, and imazethapyr is derivatized with trimethylanilinium hydroxide (TMAH). Methylated imazethapyr was analyzed by gas chromatography with a nitrogen-phosphorous dtector. The recovery of imazethapyr from soil samples are over 80%, the experimental detection limit of the method is 5ng·g -1 .

关键词： imazethapyr residue gas chromatography

Partial Characteristics of Hydrogen Production by Fermentative Hydrogen-producing Bacterial Strain B49

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High Technology Letters 2003年第3期9卷 65-70页

作者：王相晶 Ren Nanqi xiang wensheng Lin Ming Guo Wanqian Research Center of Life Science and Biotechnology Northeast Agricultural University Harbin 150030 P.R.China School of Municipal & Environmental Engineering Harbin Institute of Technology Harbin 150090 P.R.China School of Municipal & Environmental Engineering Harbin Institute of Technology Harbin 150090 P.R.China Research Center of Life Science and Biotechnology Northeast Agricultural University Harbin 150030 P.R.China

To investigate the characteristics of hydrogen production by a novel fermentative hydrogen-producing bacterial strain B49 (AF481148 in EMBL), batch experiments are conducted under different conditions. Hydrogen production has a correlation with cell growth and the consumption of glucose and soluble protein. The optimum pH for cell growth is 4.5±0.15. At acidic pH 4.0±0.15, the bacteria has the maximum accumulated hydrogen volume of 2382 ml/L culture and the maximum hydrogen evolution rate of 339.9 ml/L culture·h with 1% glucose. The optimum temperature for cell growth and hydrogen production is 35℃. In addition, fermentative hydrogen-producing bacterial strain B49 can generate hydrogen from the decomposition of other organic substrates such as wheat, soybean, corn, and potato. Moreover, it can also produce hydrogen from molasses wastewater and brewage wastewater, and hydrogen yields are 137.9 ml H 2/g COD and 49.9 ml H 2/g COD, respectively.

关键词： hydrogen production hydrogen-producing bacterial strain B49 characteristics

Improvement of Ramoplanin Production by Mutation and Medium Optimization

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Journal of Northeast Agricultural University(English Edition) 2008年第1期15卷 18-22页

作者： WU xiangjin WANG xiangjing SHI Yingzi xiang wensheng BAI Hua College of Life Sciences Northeast Agricultural University Harbin 150030 China Zhejiang Hisun Pharmaceutical Co. Ltd. Taizhou 318000 Zhejiang China

The aim of this work was to develop a mutation and selection procedure to select the high-yield strain, and this strain could alleviate the inhibition in the biosynthesis of ramoplanin caused by L-leucine. In this experiment, the strain Actinoplanes sp. C6-78-14 that produced 384.8 mg of total productivity per liter was isolated. Its ramoplanin productivity was 4.13-fold higher than that of the parent strain A ctinoplanes sp. C2-9-45. The subculture experiments showed that the hereditary character of C6-78-14 was stable. Subsequent media studies with strain Actinoplanes sp. C6-78-14 showed that adding 0.001% FeSO4· 7H2O, 0.001% CoCl2· 6H2O and 0.001% CuSO4· 5H2O to fermentation medium respectively could enhance the ramoplanin production, and the production of each group was increased by 10%, 15% and 20% compared with the control group.

关键词： ramoplanin 4-aza-leucine mutation fermentation

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色谱法在糖类化合物分析中的应用

分析化学 2001年第2期29卷 222-227页

作者：王静王睛向文胜东北农业大学生命科学与生物技术研究中心!哈尔滨150030

综述了适用气相色谱分析和高效液相色谱分析多糖的降解方法，单糖和寡糖衍生物的制备，色谱条件的选择以及色谱法在糖类分析中的应用。参考文献５８篇。

关键词：色谱法糖类化合物评述