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Table of Content

    05 February 2015, Volume 31 Issue 2
    Review and editorial
    Cultivation of Microalgae with Flue Gas: Mechanism and Application
    Du Kui, Liang Fang, Geng Yahong, Li Yeguang
    2015, 31(2):  1-9.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.001
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    Biodiesel from microalgae is considered as the only renewable biofuel that has the potential to displace traditional petroleum-derived transport fuels and meet the global demand for transport fuels. However, its more widespread use is limited by high cost of microalgal cultivation. Industrial waste gas(flue gas) contains not only a lot of CO2, but also considerable sulfur oxides(SOx) and nitrogen oxides(NOx), so utilization of flue gas for microalgal cultivation to reduce the cost of microalgal biodiesel has atracted more and more attention. In this paper, the mechanism of absorption and metablization of CO2, SO2 and NOx by microalgae and the applications of flue gas in cultivation of microalgae were reviewed. Based on the unique capacity of microalgal suspention culture system in utilization of CO2, SO2 and NOx, the idea of simultaneous lipid production, CO2 fixation, SO2 and NOx removal by microalgae was established.
    Progress on Regulation of Anther Dehiscence by Jasmonic Acid
    Guo Hang, Wang Zhimin, Tang Qinglin, Tian Shibing, Yang yang, Song Ming
    2015, 31(2):  10-17.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.002
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    Jasmonic acid(JA) is a ubiquitously occurring plant growth regulator. JA and methyl jasmonate(MeJA) play important role in plant life. JA is involved in stamen development, regulates anther dehiscence and affects plant fertility. This paper reviewed and summarized the JA biosynthesis pathway and the regulation of gene expression, and the molecular research on JA regulates plant anther development, especially anther dehiscence at the late stage. Finally it puts forward some prospects for future study.
    Research Advances on Plant Promoter
    Li Tian, Sun Jingkuan, Liu Jingtao
    2015, 31(2):  18-25.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.003
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    Plant promoters play important regulatory roles at the transcription level, to study their functions can not only reflect the expression patterns of corresponding genes, but also provide an effective way for the use of plant genetic engineering to achieve efficient expression of specific genes. In this paper, recent studies were reviewed about their structural characteristics and functions, the various types of inducible promoters related to biotic and abiotic stress were mainly introduced, and the future research directions for plant promoters were prospected.
    Advances in Studies on Genomics of Toxogenic Fungi
    Wang Yan, Liu Yang, Liu Yining
    2015, 31(2):  26-34.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.004
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    Mycotoxins, a group of toxins structurally related to secondary metabolites are produced by some fungal strains. It is mainly produced by filamentous fungi, including Trichoderma, Aspergillus, Chaetomium, Penicillium and Rhizopus, existed in agricultural products. Illustrating the genome sequence information of the pathogenic microorganisms in agricultural products, is essential to reveal fungal specific genetic traits. Until to December 2013, a total of 204 species in Ascomycetes and 62 species in Basidiomycete genomic information had been sequenced or published, genomes size from approximately 30 Mb to 40 Mb. This review here provides an overview of available genomic information of toxin-producing fungal or virulent fungus, summarizes the recent development in genome sequencing strategies and main fungal produced mycotoxin, and proposes the future direction of the comparative genomics research on fungi.
    Research Progress of Bacillus subtilis Expression System and Its Promoter Regulatory Elements
    Yu Xiaoxia, Tian Jian, Liu Xiaoqing, Wu Ningfeng
    2015, 31(2):  35-44.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.005
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    As a Gram-positive bacteria, Bacillus subtilis is an attractive host for the production of heterologous secretory proteins for several reasons:it is non-pathogenic and the capable of secreting functional extracellular proteins directly to the culture medium, a great deal of vital information concerning large scale fermentation and production technology. One of the key factors for achieving high-level expression of heterologous proteins is the use of a strong and control promoter. In present, the promoters of Bacillus subtilis can be classified into three categories:constitutive promoters, inducer-specific promoters and autoinducible promoters. This paper described the advantages and disadvantages of Bacillus subtilis expression system and the classification of promoters. At the same time, we summarized the methods of the amplification of new promoters, which provided a foundation for improving Bacillus subtilis expression systems and the industrial production of heterologous proteins .
    Research Development of Functions of Peptidoglycan Hydrolase Produced by Bacillus
    Wang Dandan, Guo Shuyuan
    2015, 31(2):  45-52.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.006
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    Over the years, peptidoglycan hydrolases are considered destructive roles, for example the phage lysis host use them to release the virus particles, some bacterias dissolve their competitors, etc. But in recent years, with the continuous research depth found:in cell growth and metabolism process, its positive roles are more important. This paper reviewed the functions of peptidoglycan hydrolase in Bacillus throughout the cell life course, including cell growth, division, motility, sporulation and spore germination, protein secretion, information transmission, innate immunity and other aspects, laid the theoretical foundation for the study of peptidoglycan hydrolases and cell metabolism in Bacillus and the other microorganism.
    The Structure-function Relationship of Vitreoscilla Haemoglobin and Its Application in Biotechnology
    Chen Yunmei, Xu Haiyang, Wang Shiming
    2015, 31(2):  53-60.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.007
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    The hemoglobin from the bacteriumVitreoscilla(VHb) is the firstmicrobial hemoglobin, which has been extensively studied with respect to its structure, biochemical functions, and the mechanisms by which itsexpression is controlled. Expressing VHb in a variety of heterologous hostsresults substantial increasesin production of a variety of useful products(antibiotics, proteins, polymersetc.), host resistance andability to degrade some harmful compounds, by promoting oxygen transport to enhance respiratory and energy metabolism. So this paper reviewed on the function and structure of VHb and the improving output of microbial metabolites, some characteristics of plants and animals and influence of the application of bioremediation capability. And we discussed about the application prospect and development space in the metabolism of plant and animal.
    Polyamine Biosynthesis Enzyme Research Progress in Two Key Genes
    Lü Huanqing, Wang Zhimin, Tang Qinglin, Tian Shibing, Wang Yongqing, Song Ming
    2015, 31(2):  61-64.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.008
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    Polyamine is an important physiological regulation substance functioning in a wide variety of biological processes, such as plant growth, development, senescence and adversity stress tolerance, which widely exist in all living organisms. S-adenosylm ethionIne synthetase gene(SAMS) and S-adenosylmethionine decarboxylase gene(SAMDC) are the two key genes in the polyamine synthesis pathway. This paper summarized the gene cloning, expression and transgenic expression regulation of SAMS and SAMDC, and other aspects also reviewed. Its application prospect was discussed in the end.
    Advance in the Expression Pattern and Regulatory Mechanism of BMP1 on Animals Folliculogenesis and Embryo Development
    Lei Xiaocan, Li Lanyu, Li Zhipeng, Cui Kuiqing, Liu Qingyou, Shi Deshun
    2015, 31(2):  65-70.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.009
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    Bonemorphogeneticprotein 1(BMP1) is a kind of metalloproteinases belongs to the astacin family, its play an important role in the process of the formation of extracellular matrix and regulating TGFβ signaling pathways. BMP1 gene expressed in the follicular development with different expression levels, and involved in the embryogenesis and development of animal embryos in some way. The research on BMP1 gene may help people from genetic point of view to clarify the mechanisms of the formation follicular and embryo development, and then may have a positive impact on animal husbandry and other fields of medicine. This review will focus on BMP1 gene expression in the process of animal embryonic development orientation, function and its mechanism involved.
    Technique
    Advances and Applications on Methodology of 16S rRNA Sequencing in Gut Microbiota Analysis
    Li Dongping, Guo Mingzhang, Xu Wentao
    2015, 31(2):  71-77.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.010
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    16S rRNA sequencing is one of the high-throughput-sequencing-based methods used in gut microbiota analysis. Almost all the bacterial species in gut microbiota can be quantified through 16S rRNA sequencing, which has made this method into the mainstream. Two issues are very important in the application of 16S rRNA sequencing:sequencing strategy and bioinformatic analysis. In this review, three aspects of the sequencing strategy, including sequencing platform, sequencing region, and data size were discussed. While on bioinformatic analysis, the advance in sequences cluster and annotation, microbiota structure analysis, key taxa screening and functional analysis were reviewed here.
    Rapid Identification of Four Kinds of Decay Fruit Moths by Real-time Fluorescence Quantitative PCR
    Wang Fengjun, Liu Shasha, Feng Junli
    2015, 31(2):  78-83.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.011
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    Laspeyresia pomonella, Euzophera pyriella Yang, Grapholitha molesta and Blank cutworm are important pets of fragrant pears and other fruits in Xinjiang, and they are also internationally important pests on inspection and quarantine. However, the identification relies mainly on morphological characters up to date. The lack of molecular detection methods greatly limits the international export trading of fragrant pears and related products. The 16S rDNA COI fragments from the four pests were cloned and sequenced. Then, specific primers and probes were designed according to the sequencing data, and the fluorescence Real-time quantification PCR detection system was established. The results demonstrates the limits of detection concentration of Laspeyresia pomonella, Euzophera pyriella Yang, Grapholitha molesta and Blank cutworm in this system are respectively 1.605×10-2,0.729×10-2,0.475×10-2 and 0.818×10-2ng/μL, meeting the demand of inspection and quarantine routine testing. To overcome the residual bodies of some insects and some pieces mixed samples, or several insects together mixed samples appeared in actual testing process, DNA extracted from single, multiple and mixed pest samples were detected and evaluated, further verifying the specificity of this method, the reliability and applicability.
    Research Report
    Screening a New Type of Schizochxtrium Strain Producing DHA Using Starch by Protoplast Fusion Technology
    Wang Di, Lu Fuping, Wang Haijun, Li Demao, Chen Shulin, Zhang Ke
    2015, 31(2):  84-90.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.012
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    Schizochxtrium B4D1 and Aspergillus niger CGMCC 3.316 as original strains, a new strain of Schizochxtrium was selected using starch for producing DHA. The protoplasts from two parents strains were obtained respectively with Lysing enzyme. The study was investigated on the effect of culture time, culture method, enzymolysis time on the protoplasts preparation. Finally, the protoplasts fusion was done under the PEG mediated condition. The optimum condition of protoplast fusion was as follows:PEG6000 concentration of 40%, fusion temperature of 30℃, fusion time of 10 min. Furthermore, the fusion rate reached 1.9% at the optimum condition. Through comparing its colony characteristics, color, morphology and purified spore culture, a new strain has been selected using starch as carbon source. RAPD verification showed that the fusant having integrated genome of B4D1 and CGMCC 3.316 expressed more genetic information from B4D1.
    Research of PKS2 Interaction with CAM4 in Arabidopsis thailana
    Qiao Xinrong, Duan Hongbin, Liu Zhuming, Zhao Fuan
    2015, 31(2):  91-96.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.013
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    Blue-light receptors phototropin (PHOT)mediate a wide set of physiological and developmental responses. As some PHOT downstream signal transduction components have been identified in Arabidopsis by genetic analyses, we have found that PKS1 of Phytochrome kinase substrate PKS family interacts with members of calmodulin (CAM), which are Ca2+ binding protein. In order to address the issue of how PKS2 interacts with CAM4, firstly cDNA full length sequences of PKS2 and CAM4 were obtained by RT-PCR technique. Then, it was confirmed that PKS2 could be interact with CAM4 by tests of yeast two-hybrid system and bimolecular fluorescence complementation. These date could contribute to enrich relation between PKS and CAM, which would provide base for further uncovering PHOT functions.
    Optimization and Performance Evaluation of the Regeneration System of the Baodi Garlic
    Fan Baoli, Wang Zhen, Ren Chunxue, Gu Zenghui, Liu Xiaoying, Wang Zhenying
    2015, 31(2):  97-102.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.014
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    The regeneration system of Baodi garlic was improved by optimizing the differentiation medium and rooting medium. The optimal differentiation medium was MS +6-BA 5.0 g/L + NAA 1.0 g/L and the best medium for rooting was MS medium without hormones. No chromosomal aberration was observed according to the results of the cytological identification and field assessment of the plant regeneration. The field features indicated the improvement of the photosynthetic efficiency of the plant regeneration.
    Cloning and Sequence Analysis of CYP83A1 Gene from Brassica oleracea var. italica
    Cao Yanglihui, Li Yong, Kong Wenwen, Li Jing
    2015, 31(2):  103-110.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.015
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    To further research the related genes involved in 4-methylsulfinybutyl-glucosinolate biosynthesis in Brassica oleracea var. italica, the key synthetase CYP83A1gene was cloned and analyzed by bioinformatics. The sequence of CYP83A1 from the previous data of broccoli transcriptome sequencing and the sequences of CYP83A1 in seven other plants (Arabidopsis thaliana, Brassica napus and the Brassica rapa subsp. Pekinesis et.al)which obtained from GenBank were alignmented with each other using NCBI blaster, and we confirmed that the homologically conserved region of CYP83A1 gene in Brassica oleracea var. italicaBoCYP83A1). Using RT-PCR method, the CDS region of BoCYP83A1 was obtained. Bioinformatic analysis indicated that the CDS region of BoCYP83A1 was 1 509 bp long which encoded a predicated a protein contains 502 amino acids. Protein analysis showed that BoCYP83A1 molecular weight was 57.47 kD, theoretical pI was 7.1 and contained two span membrane structures, and the BoCYP83A1 does not have signal peptide sequences, while possessed a P450 functional domains. Homology analysis of the deduced amino acids indicated that BoCYP83A1 had 98% similarity with Brassica napus and the Brassica rapa subsp. Pekinesis. The gene registration number of Brassica oleracea var. italica CYP83A1 gene is KM111290.
    Cloning and Expression Analysis of Actin Gene Gragment from Elsholtzia haichowensis
    Cai Shenwen, Xiong Zhiting, Liu Chen, Xu Zhongrui, Deng Songqiang
    2015, 31(2):  111-115.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.016
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    Cloning and expression analysis of Actin gene fragment from Elsholtzia haichowensis would provide foundation for the study of gene expression and regulation of heavy metal resistance related genes. Degenerate primers were designed based on the conserved sequences of the Actin genes from other plants. Total RNA was extracted from the root of E. haichowensis. An Actin gene fragment was separated by reverse transcription polymerase chain reaction(RT-PCR). The sequence analysis results revealed that Actin gene fragment from E. haichowensi contains 576 bp, encoding a protein of 192 amino acids. Homology comparison with other plants Actin gene sequences in the GenBank showed that it shared 84%-97% amino acid sequence homology with other plants. The cloned sequence was Actin gene fragment. It was named as EhACT and was registered into GenBank(accession number:AGT37260). Semi-quantitative PCR assays indicated that the expression of EhACT in root, stem and leaf of E. haichowensis was relatively stable, suggesting that EhACT can be used as the reference to analyze the gene expression in E. haichowensis.
    Cloning and Characterization of HbSnRK2.4 in Tibetan Hulless Barley(Hordeum vulgareL. var. nudum HK.f.)
    Zeng Xingquan, Wang Yulin, Xu Qijun, Yuan Hongjun , Wei Zexiu, Ni Mazhaxi
    2015, 31(2):  116-121.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.017
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    Genetic Variation Study of NGB Gene Exon 3 of Simmental Cattle Hybrid Taxon in Gansu Hexi Regions
    Liang Chunhua, Liu Xia, Sun Xuejing, Luo Yuzhu, Gao Xuejin, Du Xiaohua
    2015, 31(2):  122-128.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.018
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    This study was designed to systematically analyze the genetic polymorphism and variability of NGB gene exon 3 of 283 Simmental cattle hybrid taxon in Gansu Linze, Ganzhou, Wuwei, Jinchang, and Gaotai. DNA fragment containing the exon 3 and part of intron of NGB gene was amplified and genotyped using PCR single-strand conformational polymorphism(SSCP) method. Representative alleles were sequenced for verification of their variations in DNA sequences. The results displayed that there were 5 alleles(A, B, C, D and E) detected, combined five genotypes as AA, AB, AC, AD and AE. Among them, samples from Ganzhou, Wuwei and Jinchang were detected AA and AB genotypes, AA and AE genotypes in Gaotai and AA, AB, AC and AD genotypes in Linze. Allele A was the predominant allele and AA was the predominant genotype in five breeds. Sequence of the SSCP showed 6 mutations in the DNA fragment(75 bp C→T, 78 bp C→G, 128 bp G→A, 214 bp G→A, 232 bp C→T, and 233 bp G→A). In which, C→T at 75 bp and C→G at 78 bp were located in intron, while the other mutations were located in exon. G→A mutation at 214 bp, C→T mutation at 232 bp and G→A mutation at 233 bp were lead to Gly→Ser, Arg→Trp and Arg→Gln, respectively. Chi-square testing indicated the three gene mutation were all in Hardy-Weinberg equilibrium(P> 0.05). Population genetics analysis showed the polymorphism information content were 0.0582, 0.0196, 0.0196, 0.0161 and 0.0159 in Linze, Ganzhou, Wuwei, Jinchang, Gaotai, respectively, which was at low polymorphism(PIC <0.25).
    Cloning and Bioinformatics Analysis of the CDS of Yak AQP4 Gene
    Liu Jianfeng, Ding Yanping, Hou Boru, Wang Jianlin, Shao Baoping
    2015, 31(2):  129-134.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.019
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    A coding region sequence of yak AQP4 was cloned by molecular cloning technique. Some characters of the AQP4 gene and encoded protein sequences were predicted and analyzed by the methods of bioinformatics in the following aspects as the general physical and chemical properties, hydrophobicity, transmembrane structure and secondary structure. Results showed that the full-length of yak AQP4 contains a complete ORF(966 bp) encoding 322 amino acid. The putative molecular weight and theory isoelectric point of AQP4 gene in yak were 34.69 kD and 7.59, respectively. The protein encoded by yak AQP4 contains six transmembrane α-helices, and it is a hydrophobic protein. The second structures are mainly composed of α-helix, extended strand and random coil. Deduced amino acid sequences of AQP4 gene between yak and Bos Taurus, Ovis aries, Susscrofa and other species were high on homology and the phylogenetic distance consistent with their genetic relationship.
    Molecular Cloning and Expression Analysis of MyD88 in Roughskin Soulpin,Trachidermus fasciatus
    Bi Caihong, Zhang Qiuxia, Yu Shanshan, Chen Xuezhao, Liu Chunying, Zhu Qian
    2015, 31(2):  135-142.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.020
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    Myeloid differentiation factor 88(TfMyD88) is a key adaptor protein in the Toll-madiated signaling passway. In this study, we cloned the full-length cDNA from the roughskin soulpin, Trachidermus fasciatus. The full-length of MyD88 was 1 555 bp, which contained an open reading frame(ORF) of 867 bp encoding a polypeptide of 288 amino acids. The deduced amino acid sequence has a conserved death domain(DD) at the N-terminal and a typical Toll/interleukin-1 receptor(TIR) domain at the C-terminal. The mRNA expression patterns of TfMyD88 in healthy and LPS challenged roughskin soulpin were detected using quantitative Real-time PCR. MyD88 was expressed broadly in the blood, heart, liver, gill, intestine, skin, musle, kidney, spleen, brain, with the highest expression in the gill. The expression of TfMyD88 post challenge with LPS(lipopolysaccharide) was detected in blood, liver, gill, skin and spleen. The up-regulation of the expression levels of TfMyD88 in all the detccted tissues after chanllaged by LPS suggests that MyD88 plays an important role in roughskin soulpin defenses against bacterial infection.
    Cloning and Prokaryotic Expression of Musca domestica Thymosin Gene
    Wang Yu, Wu Gaoji, Luo Man, Peng Chuanlin, Xiu Jiangfan, Shang Xiaoli, Wu Jianwei
    2015, 31(2):  143-147.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.021
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    In order to clone and analyze the prokaryotic expression of thymosin gene from Musca domestica, the thymosin gene which was isolated from Musca domestica cDNA library, was analyzed by the bioinformatics methods in the following aspects, including general physical and chemical properties, signal peptide and subcellular localization. The expression construct pET-28a(+)-THY was preformed. The open reading frame of the thymosin gene was 384 bp that encoded a putative protein with 127 amino acids. The protein with predicted molecular weight 14.3 kD and pI of 5.22, has the conserved thymosin domain that belongs to thymosin family. The result showed that the recombinant prokaryotic expression vector pET-28a (+)-THY was successfully constructed and fusion protein was expressed in E. coli. SDS-PAGE and Western blot analysis indicated that the fusion protein that purified using Ni2+ affinity chromatography had the predicted size.
    Sexual Hybridization Breeding Selection for Good Quality Strain of Tremella fuciformis
    He Yuanchuan, Chen Shijiang, Yang Yong, He Zongyi , Zhang Deli, Chai Jiayan
    2015, 31(2):  148-152.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.022
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    During the process of Tremella fuciformis cultivation, some problems, such as low yield and long cultivation time, often are followed with different strains. The aim of this study is to mix good properties of different strain together to new strains by sexual hybridization method. Based comparison of yield, fresh ear ventral color and mature culture, yinke 1# and Tr21 were selected as parents strains with good properties. 50 single spores were isolated from fresh ear and partnered each other following single hybridization principle. 17 combinations are identified with an optical microscope examination and 7 genetic characteristics of hybrid strains are different from parent strains by Antagonism experiment. The results of agronomic traits comparison showed that cultivation period, diameter and the yield of hybrid strains GT3 are significantly different from the parents, and the production has exceeded the parent strains more than 3%. GT3 can be used as an excellent hybrid strains reserves and has the potential to develop into a new species. Sexual hybridization breeding would be effective way to select good quality strain.
    Cloning,Expression and Characterization of Acetyl Xylan Esterase from Streptomyces griseus
    Liu Weina, Liang Di, Wang Xiaoyu, Yu Wangning, Hou Lingyu, Jin Yi, Xie Xiangming
    2015, 31(2):  153-159.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.023
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    Primers were designed by putative axe annotated from the whole genome sequence of Streptomyces griseus. The axe gene was firstly cloned and linked to the prokaryotic expression vector, expressed recombinant protein induced with IPTG and purified with the Ni-NTA affinity chromatography. The results showed the cloned gene axe contained an open reading frame of 1 008 bp encoding 336 amino acid residues. The transformant harboring pET28a-Sgraxe was expressed with a protein molecular weight of 37 kD by SDS-PAGE analysis. The results indicated that the purified recombinant enzyme exhibited optimum activity at pH8.0 and 50℃, high thermal stability and strong alkali resistance range. Metal ions on recombinant enzyme activity was inhibited especially Zn2+. The analysis of recombinant enzymatic properties reveals potential application in industry.
    Isolation, Expression and Identification of Multifunctional Glucoamylase from Marine Streptomyces olivaceus FXJ7.023
    Wang Miao, Li Yuanyuan, Yue Changwu, Shao Meiyun , Lü Yuhong, Bao Yuxin, Huang Ying
    2015, 31(2):  160-166.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.024
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    SoGA, a predicted glucoamylase encoded by a full-length 1 788 bp gene consists of 595 amino acids was cloned from Streptomyces olivaceus strain FXJ7.023 genomic DNA by PCR. Heterologous expression of SoGA in Escherischia coli strains BL21(DE3) plysS was performed and a fusion protein with molecular weight of 83.22 kD was obtained. The novel recombinant glucoamylase showed multifunctional catalytic activity of hydrolyze carboxymethylcellulose and starch under different temperature and pH.
    Immobilization of Glucoamylase onto the Electrospinning PEI/PVA Composite Nanofiber Membrane via Ionic Adsorption
    Sui Chunhong, Wang Cheng, Dong Shunfu, Han Liqin
    2015, 31(2):  167-172.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.025
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    It was to study the physical and chemical properties of immobilization of glucoamylase on the PEI/PVA nanofibers via ion bonding. Polyethyleneimine(PEI)/polyvinyl alcohol(PVA) composite nanofiber membrane was prepared by electrospinning technology, and thermal crosslinking was applied to make it possess water stability. Glucoamylase was immobilized onto the electrospinning PEI/PVA composite nanofiber membrane via ion adsorption. Results showed that fourier transform infrared spectrum(FT-IR) indicated that glucoamylase can be successfully fixed on the surface of the electrospinning PEI/PVA composite nanofiber membrane. Then, the enzymatic properties of immobilized glucoamylase were identified. The results have shown that the optimum reaction temperature of immobilized glucoamylase is 65℃, 6 degrees higher than free glucoamylase, and the applicable pH range of immobilized glucoamylase is wider than free glucoamylase significantly. In addition, immobilized glucoamylase also has good thermal stability, storage stability and reusability. The use of ion adsorption can help the protein molecules easily fixed onto the nanofiber membrane, so this method processes a certain application.
    Selection,Identification and Medium Optimization of a Phosphate-solubilizing Bacterium
    Huang Daming, Li Qian, Guan Guoqiang, Zhang Zhicai, Qian Jingya, Song Qingchun
    2015, 31(2):  173-178.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.026
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    A phosphorus-dissolving bacterial strain P0417 was isolated from crop rhizosphere soil and identified by the genetic identification of 16S rDNA, measuring capability of phosphate-solubilization and optimization of medium components here. The result demonstrated that the bacterium was identified as Burkholderia cepacia. And the capacity of dissolving the phosphate was closely correlated with the pH of its culture medium, the strain showed high phosphate-dissolving ability of 791.84 μg/mL in Ca3(PO42 culture medium at glucose, 10 g/L;Ammonium oxalate, 0.5 g/L;NaCl, 1.0 g/L.
    Optimization of Bacillus sublitis JH-1 for Xylanase Production by Response Surface Analysis
    Wang Chunming, Zhong Chao, Wang Fengxue, Huang Fan, Jia Honghua, Wei Ping, Zhao Yin
    2015, 31(2):  179-186.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.027
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    Xylanase producing strain Bacillus sublitis JH-1 was screened from straw compost by Congo red transparent circlemethod. According to the single-factor experiments, response surface methodology was applied to optimize the liquid state fermentation conditions of Bacillus sublitis JH-1 for xylanase production, with the optimal conditions being listed as follows:temperature as 33℃, pH value as 6.0, maltose as 2.6%(m/V), YE as 1.2%(m/V), KH2PO4 as 1.2%(m/V). Increased 1.8 times in average as compared with preliminary culture and consistent with the maxmum predicted value.
    Optimization of Production Condition of Monascus Pigment by Monascus purpureus in Liquid State Fermentation
    Zhang Rui
    2015, 31(2):  187-195.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.028
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    In order to improve the yield of Monascus pigment and reduce the production costs, Monascus purpureus MY-03 was adopted to produce Monascus pigment in the liquid state fermentation, and the fermentation conditions were optimized by Plackett-Burman design and Box-Benhnken experimental design. The results showed that the optimum medium was composed of glucose 50 g/L, peptone 58.07 g/L, MgSO4 2 g/L, NaNO3 2 g/L, MnSO4 0.3 g/L, ZnSO4 0.1 g/L;and the optimum bottle load was 50.8 mL in a 250 mL flask, and the optimum inoculum volume was 10.0%(V/V). The color value of Monascus pigment reached to 342.24±2.88 U/mL, increased by 86.9% than that in the basal medium, when the test fungi were incubated in 150 r/min and 30℃ for 7 d.
    A Effect of Liquid Alkane Oxygen-vectors on Rhodotorula Fermentations and Lycopene Production
    Li Nana, Wu Xiaoying, Wu Zhenqiang
    2015, 31(2):  196-201.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.029
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    Addition of oxygen-vectors(n-hexane, n-dodecane, hexadecane)to fermentation medium was recognized as a method of enhancing oxygen transfer and promoting lycopene yield by Rhodotorula fermentation. n-dodecane as an oxygen-vector is the best in three kinds of liquid alkane. Experimental results show that the biomass of Rhodotorula attained 16.49 g/L and the yield of lycopene biosynthetic reached 42.32 mg/L by adding 4% n-dodecane in the 0 h, which was 26.2% and 50.17% higher than the control group, respectively.
    Kinetics of Batch Fermentation of Epothilone B by Sorangium cellulosum SoF5-76
    Ma Liyun, Gong Guoli, Wang Na
    2015, 31(2):  202-207.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.030
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    The fermentation system produced epothilone is complex, and the production strain is more difficult to operate, leading to difficulties in fermentation parameters measured, so far there is no reporting on Epothilone fermentation kinetics. In this paper, the batch fermentation kinetics of the epothilone B produced by Sorangium cellulosum SoF5-76 was studied in a 5 L fermentor. During the process, the temperature and pH were almost constant, 30℃, 7.4. When the fermentation finished, the dry cell weight, epothilone B production and residual glycerol could reach 3.00 g/L, 18.20 mg/L and 0.049 g/L. On the bases of Logistic and Luedking-Piret equation, the models of cell growth, epothilone B production and substrate consumption were established, and the non-line fitting was employed to model the kinetics of fermentation by software MATLAB. The results show that the kinetic models were in good agreement with the fermentation process.
    An Investigation of Cellular Autophagy and Oil Accumulation in the Lipomyces starkeyi
    Han Bing, Yang Qin, Cui Qinghua
    2015, 31(2):  208-216.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.031
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    It was to prove autophagy is involved in the accumulation of lipid in Lipomyces starkeyi. Detecting the expression of autophagy-related gene in different lipid accumulation level to determine whether autophagy is potentially involved in lipid accumulation; promoting autophagy by autophagy accelerator to observe whether there are differences in yeast lipid accumulation level. Results showed that the level of autophagy is low in high lipid accumulation condition compare with low condition. After treatment with autophagy accelerator, the lipid accumulation in yeast is reducing. In Lipomyces starkeyi, autophagy is negatively related to lipid accumulation, the improving of autophagy level will reduce the lipid accumulation in yeast.
    The Influence Factors of Determination of Haematobacter sp. Cellular Fatty Acids
    He Weihong, An Mingli, Chen Guocan, Jia Bin, Wu Xiaolei, Wang Yanan
    2015, 31(2):  217-221.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.032
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    Cellular fatty acid composition of strain HNMC11807, a novel species belong to Haematobacter sp., were detected by used Sherlock MIS system of MIDI company USA. The influence by different culture methods and cells growth conditions on cells fatty acid composition were explored in this article. Based on the bacterial cells cultured in varied medium such as in different medium components and in solid or liquid medium, the cellular fatty acid composition were respectively analysed and compared. On the same time, two standard systems in this instrument, conventional and fast, were used to run samples, and the detect results of the cellular fatty acid composition were compared respectively. As the results, the cells fatty acid composition was closely related with culture condition and a significant differences. The cellular fatty acid composition must be carried out in specified and unified culture conditions when this system was used in microbial classification.
    Prokaryotic Expression and Purification of Zinc Finger Domain of Human Pokemon Protein
    Liu Li, Li Yueting, Zhang Mengmeng, Yang Yutao, Xu Zhiqing
    2015, 31(2):  222-227.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.033
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    It was to express human Zinc finger domain of Pokemon gene in prokaryotic cells and purify the GST-Zinc finger fusion protein. The segment of zinc finger domain of Pokemon gene was amplified by PCR and cloned into prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1-Zinc finger was transformed into E.coli BL21(DE3)and exogenous protein was induced by IPTG. After purification using MagneGST particles, the GST-Zinc finger fusion protein was further identified by Western blot. Results showed that the recombinant plasmid pGEX-4T-1-Zinc finger was constructed successfully. When BL21(DE3) cells transformed with pGEX-4T-1-Zinc finger were cultured at 30℃ and induced with 0.2 mmol/L IPTG, GST-Zinc finger protein was obtained in a large quantity in supernatant. The purified GST-Zinc finger was further identified specifically by Pokemon antibody. Therefore, it proved that GST-Zinc finger fusion protein was successfully expressed and purified, and could be used for further study of the function of Pokemon.
    Growth Influence of Allicin and Its Prodrug on Esophageal Cancer Cells
    Yang Jinbu, Chang Quan'e, Gou Ping, Wei Hongyan
    2015, 31(2):  228-232.  doi:10.13560/j.cnki.biotech.bull.1985.2015.02.034
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    We observed the cancer cell morphology, determined lactate dehydrogenase(LDH) activity and DNA degradation after allicin and its prodrug(alliin + alliinase) treated human esophageal cancer Eca9706 cells. The results showed that the cancer cells became more circular and smaller, the number of living cells decreased, the non-adherent cells and cell debris increased. DNA-Ladder revealed to exist DNA ladder line, which indicated DNA molecule degraded. Cancer cells’ LDH activity treated with allicin and its prodrug was significantly higher than the control group. LDH activity had a time and dose dependent manner with treated concentration increased and time extended. Eca9706 cells’ LDH activity by the prodrug treated was higher than allicin, this confirmed prodrug could cause great damage on cell membrane.