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Biotechnology Letters

Yoshiaki Tamaru, Kiwamu Umezawa, Makoto Yoshida
OBJECTIVES: The aim of the study was to obtain information about the enzymatic properties of aryl-alcohol oxidase from the plant saprophytic basidiomycete Coprinopsis cinerea (rCcAAO), which is classified into the auxiliary activities family 3 subfamily 2 (AA3_2). RESULTS: The gene encoding AAO from the plant saprophytic basidiomycete Coprinopsis cinerea (CcAAO) was cloned, and the recombinant CcAAO (rCcAAO) was heterologously expressed in the methylotrophic yeast Pichia pastoris...
May 22, 2018: Biotechnology Letters
Olufemi Emmanuel Bankefa, Meiyu Wang, Taicheng Zhu, Yin Li
OBJECTIVES: To systematically explore the effects of overexpressing Hac1p homologues from different sources on protein secretion in Pichia pastoris system. RESULTS: Effects of Hac1p homologues encompassing P. pastoris (PpHac1p), S. cerevisiae (ScHac1p), Trichoderma reesei (TrHac1p) and Homo sapiens (HsXbp1), on secretion of three reporter proteins-β-galactosidase, β-mannanase and glucose oxidase were investigated. No individual Hac1p was optimal for all the enzymes...
May 21, 2018: Biotechnology Letters
Ningning Sun, Yuanchao Qian, Weiwei Wang, Yaohua Zhong, Meixue Dai
OBJECTIVE: Improve the hydrolysis efficiency of the Trichoderma reesei cellulase system by heterologously expressing cellobiohydrolase Cel7A (Te-Cel7A) from the thermophilic fungus Talaromyces emersonii. RESULTS: Te-Cel7A was expressed in T. reesei under control of the cdna1 promoter and the generated transformant QTC14 could successfully secrete Te-Cel7A into the supernatant using glucose as carbon source. The recombinant Te-Cel7A had a temperature optimum at 65 °C and an optimal pH of 5, which were similar to those from the native host...
May 19, 2018: Biotechnology Letters
Christopher P Wallis, Aleksandra Filipovska, Oliver Rackham
OBJECTIVE: To increase the reporter repertoire of the yeast three-hybrid system and introduce the option of negative selection. RESULTS: Two new versions of the yeast three-hybrid system were made by modifying the MS2 coat RNA-binding protein and fusing it to the Gal4 DNA-binding protein. This allows the use of Gal4 inducible reporters to measure RNA-protein interactions. We introduced two mutations, V29I and N55K into the tandem MS2 dimer and an 11 amino acid deletion to increase RNA-protein affinity and inhibit capsid formation...
May 18, 2018: Biotechnology Letters
Shan Li, Lingyan Li, Xiangfeng Xiong, Xiuling Ji, Yunlin Wei, Lianbing Lin, Qi Zhang
OBJECTIVE: This study was aimed at cloning and characterizing a novel malic enzyme (ME) gene of Mortierella isabellina M6-22 and identifying its relation with lipid accumulation. METHODS: Mime2 was cloned from strain M6-22. Plasmid pET32aMIME2 was constructed to express ME of MIME2 in Escherichia coli BL21. After purification, the optimal pH and temperature of MIME2, as well as Km and Vmax for NADP+ were determined. The effects of EDTA or metal ions (Mn2+ , Mg2+ , Co2+ , Cu2+ , Ca2+ , or Zn2+ ) on the enzymatic activity of MIME2 were evaluated...
May 18, 2018: Biotechnology Letters
Saadet Alpdağtaş, Sevil Yücel, Handan Açelya Kapkaç, Siqing Liu, Barış Binay
OBJECTIVES: To identify a robust NADP+ dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929 (LbFDH) with unique biochemical properties. RESULTS: A new NADP+ dependent formate dehydrogenase gene (fdh) was cloned from genomic DNA of L. buchneri NRRL B-30929. The recombinant construct was expressed in Escherichia coli BL21(DE3) with 6 × histidine at the C-terminus and the purified protein obtained as a single band of approx. 44 kDa on SDS-PAGE and 90 kDa on native-PAGE...
May 18, 2018: Biotechnology Letters
Weiyu Liu, Yong Cao, Yadi Guan, Changqing Zheng
OBJECTIVES: To investigate the functional roles of bone marrow stromal cell antigen 2 (BST2) in gastric cancer (GC) cells and its implications in the development of GC patients. RESULTS: BST2 was frequently overexpressed in GC tissues compared with the adjacent non-tumorous tissues, and high BST2 expression was correlated with tumor stage and lymphatic metastasis. Furthermore, in vitro experiments demonstrated that knockdown of BST2 by siRNA inhibited cell proliferation, induced apoptosis and repressed cell motility in GC cells...
May 17, 2018: Biotechnology Letters
Hoda Sabati, Hossein Motamedi
OBJECTIVE: Water in oil emulsions increase oil processing costs and cause damage to refinery equipment which necessitates demulsification. Since chemical demulsifiers cause environmental pollution, biodemulsifiers have been paid more attention. This study aims to identify biodemulsifier-producing bacteria from petroleum contaminated environments. RESULTS: As a result, several biodemulsifier producing strains were found that Stenotrophomonas sp. strain HS7 (accession number: MF445088) which produced a cell associated biodemulsifier showed the highest demulsifying ratio, 98...
May 15, 2018: Biotechnology Letters
Shiwei Zhang, Qiding Zhong, Daobing Wang, Zhanbin Huang, Guohui Li
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May 14, 2018: Biotechnology Letters
Hye Joung Kim, Kyoung-Woon Kim, Yong-Rim Kwon, Bo-Mi Kim, Yoo-Jin Kim
OBJECTIVE: In order to identify specific mesenchymal stromal cell (MSC) populations with enhanced therapeutic efficacy, we evaluated the functional changes associated with the stable expression of CD200, which is associated with immune regulatory function and osteogenic differentiation, in human bone marrow-derived MSCs (CD200/MSCs). RESULTS: We detected significantly greater osteogenesis and chondrogenesis in CD200/MSCs than in mock-transfected MSCs. In addition, the immune regulatory function of MSCs in mixed lymphocyte reactions was enhanced by CD200 gene transfection...
May 9, 2018: Biotechnology Letters
Jinhui Huang, Jianxin Zhou, Guangming Zeng, Yanling Gu, Yi Hu, Bi Tang, Yahui Shi, Lixiu Shi
OBJECTIVE: A modified method was used for cell entrapped beads (CEBs) preparation and two aeration intensities (low and high aeration intensity) was supplied as factors to investigate the change of quorum quenching performance for membrane biofouling in membrane bioreactor (MBR). RESULTS: Dehydrogenase activity and growth trend of activated sludge were improved at high aeration intensity. Compared with C-MBR (with vacant beads), QQ-MBR (with CEBs) had more stable quorum quenching activity and longer application time at high aeration intensity, in which the proteins and polysaccharides were reduced by 15 and 20%, respectively...
May 7, 2018: Biotechnology Letters
Xue-Gang Xu, Lin Gong, Tian-Ling Jiang, Yuan-Hong Li, Xing-Hua Gao, Haishan Tian, Hong-Duo Chen
OBJECTIVES: To explore potential effects of recombinant human fibroblast growth factor 20 (rhFGF20) in the growth of cultured mouse vibrissal follicles. RESULTS: The growth of cultured mouse vibrissal follicles was significantly induced by rhFGF20 in a dose dependent pattern in the in vitro vibrissal follicle organ culture model. However, too high concentration of rhFGF20 could inhibit the growth of vibrissal follicles. We further demonstrated that rhFGF20 stimulated the proliferation of hair matrix cells and activated Wnt/β-catenin signaling pathway...
April 30, 2018: Biotechnology Letters
Hasan Mollanoori, Shahram Teimourian
Gene therapy is based on the principle of the genetic manipulation of DNA or RNA for treating and preventing human diseases. The clustered regularly interspaced short palindromic repeats/CRISPR associated nuclease9 (CRISPR/Cas9) system, derived from the acquired immune system in bacteria and archaea, has provided a new tool for accurate manipulation of genomic sequence to attain a therapeutic result. The advantage of CRISPR which made it an easy and flexible tool for diverse genome editing purposes is that a single protein (Cas9) complex with 2 short RNA sequences, function as a site-specific endonuclease...
April 28, 2018: Biotechnology Letters
Anurag S Rathore, Devashish Kumar, Nikhil Kateja
Over the last several decades, researchers have time and again proposed use of non-chromatographic methods for processing of biotherapeutic products. However, chromatography continues to be the backbone of downstream processing, particularly at process scale. There are many reasons for this, critical ones being the unparalleled scalability, robustness, and selectivity that process chromatography offers over its peers. It is no surprise then that process chromatography has been a topic of major developments in resin matrix, ligand chemistry, modalities, high throughput process development, process modelling, and approaches for control...
April 27, 2018: Biotechnology Letters
Heng Li, Jing Li, Ruinan Jin, Wei Chen, Chaoning Liang, Jieyuan Wu, Jian-Ming Jin, Shuang-Yan Tang
OBJECTIVES: To improve the quality of mutagenesis libraries in directed evolution strategy. RESULTS: In the process of library transformation, transformants which have been shown to take up more than one plasmid might constitute more than 20% of the constructed library, thereby extensively impairing the quality of the library. We propose a practical transformation method to prevent the occurrence of multiple-plasmid transformants while maintaining high transformation efficiency...
April 26, 2018: Biotechnology Letters
Vincent Wiegmann, Cristina Bernal Martinez, Frank Baganz
OBJECTIVES: Establish a method to indirectly measure evaporation in microwell-based cell culture systems and show that the proposed method allows compensating for liquid losses in fed-batch processes. RESULTS: A correlation between evaporation and the concentration of Na+ was found (R2  = 0.95) when using the 24-well-based miniature bioreactor system (micro-Matrix) for a batch culture with GS-CHO. Based on these results, a method was developed to counteract evaporation with periodic water additions based on measurements of the Na+ concentration...
April 24, 2018: Biotechnology Letters
Yingying Zhang, Bin Tang, Guocheng Du
OBJECTIVE: To investigate the transcriptional regulation of cellobiose synthase (CBS) in Rhizopus stolonifer. RESULTS: Transcription factor XYR1 was identified as responsible for the activation of cbs. In comparison with wild-type R. stolonifer, the deletion of XYR1 resulted in transcriptional down-regulation of cbs by approximately 40%, while XYR1 over-expression increased cbs transcription up to 175%. The highest FPA activity (1.8 IU/ml) was obtained in the XYR1-overexpressing strain OExyr1 cultivated in a 2% (m/V) glucose media, corresponding to a 96% increase compared with that of the parent strain (0...
April 21, 2018: Biotechnology Letters
Jobina Rajkumari, Subhomoi Borkotoky, Ayaluru Murali, Kitlangki Suchiang, Saswat Kumar Mohanty, Siddhardha Busi
OBJECTIVE: Anti-quorum sensing and anti-biofilm efficacy of Cinnamic acid against Pseudomonas aeruginosa was comparatively assessed with respect to potent quorum sensing inhibitor, Baicalein. RESULTS: At sub-lethal concentration, Cinnamic acid effectively inhibited both the production of the QS-dependent virulence factors and biofilm formation in P. aeruginosa without affecting the viability of the bacterium. The phytocompound interfered with the initial attachment of planktonic cells to the substratum thereby causing reduction in biofilm development...
April 21, 2018: Biotechnology Letters
Hong-Gang Duan, Fang Ji, Chun-Quan Zheng, Jing Li, Jing Wang
OBJECTIVES: To explore therapeutic effects of conditioned medium from human umbilical cord mesenchymal stem cells (hUC-MSCs) on nasal mucosa radiation damage both in vivo and in vitro. RESULTS: The mucus cilia clearance time (7 and 30 days), degree of mucosal edema (7, 30, 90 and 180 days), cilia coverage (180 days) of concentrated conditioned medium group improved compared with radiotherapy control group. The proliferation and migration abilities of irradiated and non-irradiated nasal epithelial cells significantly increased after culture in bronchial epithelial cell growth medium (BEGM) containing 10% conditioned medium of hUC-MSCs compared to cells cultured in BEGM alone...
April 17, 2018: Biotechnology Letters
Jiajia Zhang, Chuan Wu, Wei Wang, Wei Wang, Dongzhi Wei
OBJECTIVES: To develop a versatile Trichoderma reesei (teleomorph Hypocrea jecorina) expression system for the high-purity production of heterologous proteins. RESULTS: The versatile T. reesei expression system is based on xyn1 and xyn2 promoters, A824V transition in XYRI, and a bicomponent carbon source strategy. Red fluorescent protein gene rfp and alkaline endoglucanase EGV gene egv3 from Humicola insolens were used as reporter genes to test our versatile expression system CONCLUSIONS: The versatile T...
April 11, 2018: Biotechnology Letters
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