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Isolation and molecular identification of cellulolytic bacteria from Dig Rostam hot spring and study of their cellulase activity

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BIOCELL 2020年第1期44卷 63-71页

作者： Sareh HAJIABADI Mansour MASHREGHI Ahmad Reza BAHRAMI Kiarash GHAZVINI Maryam M.MATIN Department of Biology Faculty of ScienceFerdowsi University of MashhadMashhadIran Novel Diagnostics and Therapeutics Research Group Institute of BiotechnologyFerdowsi University of MashhadMashhadIran Microbiology Research Center&Department of Microbiology and Virology Ghaem Medical CenterMashhad University of Medical SciencesMashhadIran

Cellulose is the main structural component of lignocellulosic wastes that can be converted to sugars and biofuels by *** to wide applications of this enzyme in various industries around the world,cellulase is considered as the third industrial *** ability of thermophilic bacteria in the production of heat-stable cellulases has made them valuable tools in *** aim of this study was isolation and molecular identification of cellulolytic thermophile bacteria from Dig Rostam hot spring and investigating their cellulase *** were taken from water and sediments of this hot spring,and cellulolytic bacteria were enriched in media containing cellulose as the only carbon *** bacteria were incubated at 60℃,and single colonies were then isolated on solid *** red assay was used as a quick test for the qualitative screening of cellulase *** to these qualitative results,four colonies named CDB1,CDB2,CDB3,and CDB4 were isolated,and their growth curve and some other characteristics were determined by biochemical ***,endoglucanase,exoglucanase,and FPase activities of the isolates were investigated *** indicated that CDB1 exhibited the highest endoglucanase(0.096 U/mL)and exoglucanase(0.156 U/mL)activities among other isolates.16S rDNA partial sequencing indicated that CDB1 had 99%similarity to the genus Anoxybacillus,and the other isolates showed the highest similarity to the genus *** cellulase gene of CDB1 isolate with the highest cellulase activity was also cloned,and its sequence is reported for the first *** studies on this thermophilic enzyme might be useful for industrial applications.

关键词： Anoxybacillus Cellulase enzyme activity assay Thermophilic bacteria 16S rDNA

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A Method for Determination of Hexokinase activity by RP-HPLC

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Wuhan University Journal of Natural Sciences 2011年第6期16卷 535-540页

作者： GUAN Yuxia WANG Jing SUN Jiaxian College of Biological Science and Technology FuzhouUniversity Fuzhou 350002 Fujian China

A method was established to determine the activity of hexokinase by reverse-phase high-performance liquid chromatography （RP-HPLC）. The activity of hexokinase was determined by detecting the amount of ADP in the reaction that converts glucose into glucose-6-phosphate. The separation degree of ATP and ADP was as good as 3.46 （separation degree^l.5）. In the range of 0.01-0.40 mmol/L, there was a good linear relationship between concentration of ADP and its peak areas （the correlation coefficient was 0.999）. The relative standard deviation（RSD） of intraday was 1.43%-1.46%, that of interday was 2.12%-2.15%. At 30℃ （optimal temperature） and pH7.0 （optimal pH）, the recovery rate of hexokinase was 95.3%-97.6%. The results showed that the method had good precision and high recovery rate, and the enzyme activity was determined through only one-step reaction. To some extent, this method can reduce the cost.

关键词： reverse-phase high-performance liquid chromatog-raphy(RP-HPLC) hexokinase enzyme activity assay

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Cloning and analysis of a Y-tocopherol methyltransf erase gene from Brassica oleracea and the function of its recombinant protein

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Progress in Natural Science:Materials International 2003年第7期 33-38页

作者： OUYANG Qing, FAN Chuntao, SUN Hui, ZHANG Yuman, BAI Shuangyi, CAI Wenqi (1. Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China 2. College of Life Sciences, Beijing Normal University, Beijing 100875, China 3. Department of Landscape Architecture, China Agricultural University, Beijing 100094, China)

The full-length cDNA (BoTMT) of γ-TMT is obtained from Brassica oleracea by 3'- and 5'-RACE methods. The 1265 bp cDNA contains an open reading frame of 1044 bp, which encodes a protein of 347 amino acids with a predicted chloroplast transit peptide and two S-adenosylmethionine (SAM)-binding domains. Sequence analysis shows that the deduced protein shares 41. 8% -86. 5% similarity to known γ-TMTs. Semi-quantitative RT-PCR reveals that BoTMT is expressed preferentially in flowers and leaves of B. oleracea . The recombinant γ-TMT protein is obtained by cloning its encoding region into the prokaryotic expression vector pET30a. The protein expressed in E. coli accounts for 22 % of total bacterial protein. The enzyme activity assay indicates that the recombinant protein has relatively high activity to convert γ-tocopherol to α-tocopherol in vitro.

关键词： : Brassica oleracea gamma-tocopherol methyltransferase gene and expression enzyme activity assay tocopherol.

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Correlation analysis of lignin accumulation and expression of key genes involved in lignin biosynthesis of ramie(Boehmeria nivea)

Correlation analysis of lignin accumulation and expression o...

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第十九届中国作物学会学术年会

作者：唐映红 Fang Liu Hucheng Xing Kaiquan Mao Guo Chen Qingquan Guo 陈建荣 College of Life and Environmental Sciences Hunan University of Arts and Science College of Biological and Environmental Engineering Changsha University College of Agriculture Hunan Agricultural University

The phloem of the stem of ramie(Boehmeria nivea) is an important source of natural fiber for the textile industry. However, the lignin content in the phloem affects the quality of ramie phloem fiber. In this study, the lignin content and related key gene expression levels were analyzed in the phloem and xylem at different developmental *** results showed that the relative expression levels of lignin synthesis-related key genes in the xylem and phloem of the stem gradually decreased from the fast-growing period to the late maturation period, but the corresponding lignin content increased significantly. However, the relative expression levels of a few genes were the highest during the maturation period. During all three periods, the lignin content in ramie stems was positively correlated with the expression of genes, including PAL, C4 H and 4 CL1 in the phenylpropanoid pathway, F5 H and CCoAOMT in the lignin-specific synthetic pathway, and CAD in the downstream pathway of lignin synthesis, but the lignin content was negatively correlated with the expression of genes including 4 CL3 in the phenylpropanoid pathway and UDP-GT in the shunt pathway of lignin monomer synthesis. The ramie 4 CL3 recombinant protein prefers cinnamic acid as a substrate during catalysis, and it negatively regulates lignin synthesis. It is speculated that ramie 4 CL3 is mainly involved in the synthesis of ramie flavonoid compounds, and that 4 CL1 is mainly involved in lignin synthesis.

关键词： ramie(Boehmeria nivea) lignin key gene expression correlation analysis enzyme activity assay

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