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BMC Biotechnology

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https://www.readbyqxmd.com/read/28407739/construction-of-a-novel-anaerobic-pathway-in-escherichia-coli-for-propionate-production
#1
Jing Li, Xinna Zhu, Jing Chen, Dongdong Zhao, Xueli Zhang, Changhao Bi
BACKGROUND: Propionate is widely used as an important preservative and important chemical intermediate for synthesis of cellulose fibers, herbicides, perfumes and pharmaceuticals. Biosynthetic propionate has mainly been produced by Propionibacterium, which has various limitations for industrial application. RESULTS: In this study, we engineered E. coli by combining reduced TCA cycle with the native sleeping beauty mutase (Sbm) cycle to construct a redox balanced and energy viable fermentation pathway for anaerobic propionate production...
April 14, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28399854/improved-n-butanol-production-via-co-expression-of-membrane-targeted-tilapia-metallothionein-and-the-clostridial-metabolic-pathway-in-escherichia-coli
#2
Wei-Chih Chin, Kuo-Hsing Lin, Chun-Chi Liu, Kenji Tsuge, Chieh-Chen Huang
BACKGROUND: N-Butanol has favorable characteristics for use as either an alternative fuel or platform chemical. Bio-based n-butanol production using microbes is an emerging technology that requires further development. Although bio-industrial microbes such as Escherichia coli have been engineered to produce n-butanol, reactive oxygen species (ROS)-mediated toxicity may limit productivity. Previously, we show that outer-membrane-targeted tilapia metallothionein (OmpC-TMT) is more effective as an ROS scavenger than human and mouse metallothioneins to reduce oxidative stress in the host cell...
April 11, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28399848/distinctive-molecular-and-biochemical-characteristics-of-a-glycoside-hydrolase-family-20-%C3%AE-n-acetylglucosaminidase-and-salt-tolerance
#3
Junpei Zhou, Zhifeng Song, Rui Zhang, Rui Liu, Qian Wu, Junjun Li, Xianghua Tang, Bo Xu, Junmei Ding, Nanyu Han, Zunxi Huang
BACKGROUND: Enzymatic degradation of chitin has attracted substantial attention because chitin is an abundant renewable natural resource, second only to lignocellulose, and because of the promising applications of N-acetylglucosamine in the bioethanol, food and pharmaceutical industries. However, the low activity and poor tolerance to salts and N-acetylglucosamine of most reported β-N-acetylglucosaminidases limit their applications. Mining for novel enzymes from new microorganisms is one way to address this problem...
April 11, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28356086/characterization-of-calcineurin-from-cryptococcus-humicola-and-the-application-of-calcineurin-in-aluminum-tolerance
#4
Lei Zhang, Jing-Jing Zhang, Shuai Liu, Hong-Juan Nian, Li-Mei Chen
BACKGROUND: Calcineurin (CaN) is a Ca(2+)- and calmodulin (CaM)-dependent serine/threonine phosphatase. Previous studies have found that CaN is involved in the regulation of the stress responses. RESULTS: In this study, the growth of Cryptococcus humicola was inhibited by the CaN inhibitor tacrolimus (FK506) under aluminum (Al) stress. The expression of CNA encoding a catalytic subunit A (CNA) and its interaction with CaM were upregulated when the concentration of Al was increased...
March 29, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28347305/abnormal-gene-expression-in-regular-and-aggregated-somatic-cell-nuclear-transfer-placentas
#5
Bo-Woong Sim, Chae-Won Park, Myung-Hwa Kang, Kwan-Sik Min
BACKGROUND: Placental defects in somatic cell nuclear transfer (SCNT) are a major cause of complications during pregnancy. One of the most critical factors for the success of SCNT is the successful epigenetic reprogramming of donor cells. Recently, it was reported that the placental weight in mice cloned with the aggregated SCNT method was significantly reduced. Here, we examine the profile of abnormal gene expression using microarray technology in both regular SCNT and aggregated SCNT placentas as well as in vivo fertilization placentas...
March 27, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28340620/stable-high-level-expression-of-factor-viii-in-chinese-hamster-ovary-cells-in-improved-elongation-factor-1-alpha-based-system
#6
Nadezhda A Orlova, Sergey V Kovnir, Alexandre G Gabibov, Ivan I Vorobiev
BACKGROUND: Recombinant factor VIII (FVIII), used for haemophilia A therapy, is one of the most challenging among the therapeutic proteins produced in heterologous expression systems. Deletion variant of FVIII, in which the entire domain B is replaced by a short linker peptide, was approved for medical use. Efficacy and safety of this FVIII deletion variant are similar to full-length FVIII preparations while the level of production in CHO cells is substantially higher. Typical levels of productivity for CHO cell lines producing deletion variant FVIII-BDD SQ, described elsewhere, are 0...
March 24, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28302113/a-restriction-free-method-for-gene-reconstitution-using-two-single-primer-pcrs-in-parallel-to-generate-compatible-cohesive-ends
#7
Fanli Zeng, Zhimin Hao, Pan Li, Yanan Meng, Jingao Dong, Yibin Lin
BACKGROUND: Restriction-free (RF) cloning, a PCR-based method for the creation of custom DNA plasmids, allows for the insertion of any sequence into any plasmid vector at any desired position, independent of restriction sites and/or ligation. Here, we describe a simple and fast method for performing gene reconstitution by modified RF cloning. RESULTS: Double-stranded inserts and acceptors were first amplified by regular PCR. The amplified fragments were then used as the templates in two separate linear amplification reactions containing either forward or reverse primer to generate two single-strand reverse-complement counterparts, which could anneal to each other...
March 17, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28302170/identification-of-residues-important-for-the-activity-of-aldehyde-deformylating-oxygenase-through-investigation-into-the-structure-activity-relationship
#8
Qing Wang, Luyao Bao, Chenjun Jia, Mei Li, Jian-Jun Li, Xuefeng Lu
BACKGROUND: Aldehyde-deformylating oxygenase (ADO) is a key enzyme involved in the biosynthetic pathway of fatty alk(a/e)nes in cyanobacteria. However, cADO (cyanobacterial ADO) showed extreme low activity with the k cat value below 1 min(-1), which would limit its application in biofuel production. To identify the activity related key residues of cADO is urgently required. RESULTS: The amino acid residues which might affect cADO activity were identified based on the crystal structures and sequence alignment of cADOs, including the residues close to the di-iron center (Tyr39, Arg62, Gln110, Tyr122, Asp143 of cADO-1593), the protein surface (Trp 178 of cADO-1593), and those involved in two important hydrogen bonds (Gln49, Asn123 of cADO-1593, and Asp49, Asn123 of cADO-sll0208) and in the oligopeptide whose conformation changed in the absence of the di-iron center (Leu146, Asn149, Phe150 of cADO-1593, and Thr146, Leu148, Tyr150 of cADO-sll0208)...
March 16, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28298196/microbial-detoxification-of-eleven-food-and-feed-contaminating-trichothecene-mycotoxins
#9
Rafiq Ahad, Ting Zhou, Dion Lepp, K Peter Pauls
BACKGROUND: Contamination of agricultural commodities with multiple trichothecene mycotoxins, produced by toxigenic Fusarium species, is a food safety issue, which greatly affects grain production and marketing worldwide. Importantly, exposure to multiple trichothecenes may increase toxicity in animals due to their synergistic and/or additive effects. To address the problem this study aimed to achieve a novel biological trait capable of detoxifying various food and feed contaminating trichothecenes under aerobic and anaerobic conditions and wide range of temperatures...
March 15, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28292294/highly-efficient-mesophyll-protoplast-isolation-and-peg-mediated-transient-gene-expression-for-rapid-and-large-scale-gene-characterization-in-cassava-manihot-esculenta-crantz
#10
Jun-Zheng Wu, Qin Liu, Xiao-Shan Geng, Kai-Mian Li, Li-Juan Luo, Jin-Ping Liu
BACKGROUND: Cassava (Manihot esculenta Crantz) is a major crop extensively cultivated in the tropics as both an important source of calories and a promising source for biofuel production. Although stable gene expression have been used for transgenic breeding and gene function study, a quick, easy and large-scale transformation platform has been in urgent need for gene functional characterization, especially after the cassava full genome was sequenced. METHODS: Fully expanded leaves from in vitro plantlets of Manihot esculenta were used to optimize the concentrations of cellulase R-10 and macerozyme R-10 for obtaining protoplasts with the highest yield and viability...
March 14, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28288608/the-role-of-community-engagement-in-the-adoption-of-new-agricultural-biotechnologies-by-farmers-the-case-of-the-africa-harvest-tissue-culture-banana-in-kenya
#11
Sunita V S Bandewar, Florence Wambugu, Emma Richardson, James V Lavery
BACKGROUND: The tissue culture banana (TCB) is a biotechnological agricultural innovation that has been adopted widely in commercial banana production. In 2003, Africa Harvest Biotech Foundation International (AH) initiated a TCB program that was explicitly developed for smallholder farmers in Kenya to help them adopt the TCB as a scalable agricultural business opportunity. At the heart of the challenge of encouraging more widespread adoption of the TCB is the question: what is the best way to introduce the TCB technology, and all its attendant practices and opportunities, to smallholder farmers...
March 13, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28270203/the-effects-of-torc-signal-interference-on-lipogenesis-in-the-oleaginous-yeast-trichosporon-oleaginosus
#12
Felix Bracharz, Veronika Redai, Kathrin Bach, Farah Qoura, Thomas Brück
BACKGROUND: Oleaginous organisms are a promising, renewable source of single cell oil. Lipid accumulation is mainly induced by limitation of nutrients such as nitrogen, phosphorus or sulfur. The oleaginous yeast Trichosporon oleaginosus accumulates up to 70% w/w lipid under nitrogen stress, while cultivation in non-limiting media only yields 9% w/w lipid. Uncoupling growth from lipid accumulation is key for the industrial process applicability of oleaginous yeasts. This study evaluates the effects of rapamycin on TOR specific signaling pathways associated with lipogenesis in Trichosporon oleaginosus for the first time...
March 7, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28270131/identification-of-two-transcription-factors-activating-the-expression-of-osxip-in-rice-defence-response
#13
Yihua Zhan, Xiangyu Sun, Guozeng Rong, Chunxiao Hou, Yingying Huang, Dean Jiang, Xiaoyan Weng
BACKGROUND: Xylanase inhibitors have been confirmed to be involved in plant defence. OsXIP is a XIP-type rice xylanase inhibitor, yet its transcriptional regulation remains unknown. RESULTS: Herbivore infestation, wounding and methyl jasmonate (MeJA) treatment enhanced mRNA levels and protein levels of OsXIP. By analyzing different 5' deletion mutants of OsXIP promoter exposed to rice brown planthopper Nilaparvata lugens stress, a 562 bp region (-1451 - -889) was finally identified as the key sequence for the herbivores stress response...
March 7, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28264682/an-engineered-food-grade-lactococcus-lactis-strain-for-production-and-delivery-of-heat-labile-enterotoxin-b-subunit-to-mucosal-sites
#14
Nan Sun, Rongguang Zhang, Guangcai Duan, Xiaoyan Peng, Chen Wang, Qingtang Fan, Shuaiyin Chen, Yuanlin Xi
BACKGROUND: Recent researches have been focusing on mucosal immune adjuvants, which play the key roles in mucosal immunization and have become the limitation for non-injected vaccine development. Escherichia coli heat-labile enterotoxin B subunit (LTB) was regarded as a promising mucosal adjuvant for its nontoxicity and potent activity. LTB preparation issues have always been recurring, in part owing to that the recombinant LTB expressed by E. coli does not act as its native form. RESULTS: We constructed an engineered Lactococcus lactis strain using a food-grade expression system...
March 6, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28245848/a-streamlined-method-for-the-design-and-cloning-of-shrnas-into-an-optimized-dox-inducible-lentiviral-vector
#15
Sander B Frank, Veronique V Schulz, Cindy K Miranti
BACKGROUND: Short hairpin RNA (shRNA) is an established and effective tool for stable knock down of gene expression. Lentiviral vectors can be used to deliver shRNAs, thereby providing the ability to infect most mammalian cell types with high efficiency, regardless of proliferation state. Furthermore, the use of inducible promoters to drive shRNA expression allows for more thorough investigations into the specific timing of gene function in a variety of cellular processes. Moreover, inducible knockdown allows the investigation of genes that would be lethal or otherwise poorly tolerated if constitutively knocked down...
February 28, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28245836/functional-expression-of-a-novel-%C3%AE-amylase-from-antarctic-psychrotolerant-fungus-for-baking-industry-and-its-magnetic-immobilization
#16
Lei He, Youzhi Mao, Lujia Zhang, Hualei Wang, Siti Aisyah Alias, Bei Gao, Dongzhi Wei
BACKGROUND: α-Amylase plays a pivotal role in a broad range of industrial processes. To meet increasing demands of biocatalytic tasks, considerable efforts have been made to isolate enzymes produced by extremophiles. However, the relevant data of α-amylases from cold-adapted fungi are still insufficient. In addition, bread quality presents a particular interest due to its high consummation. Thus developing amylases to improve textural properties could combine health benefits with good sensory properties...
February 28, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28245818/constitutive-expression-of-active-microbial-transglutaminase-in-escherichia-coli-and-comparative-characterization-to-a-known-variant
#17
Gabe Javitt, Zohar Ben-Barak-Zelas, Moran Jerabek-Willemsen, Ayelet Fishman
BACKGROUND: Microbial transglutaminase (mTG) is a robust enzyme catalyzing the formation of an isopeptide bond between glutamine and lysine residues. It has found use in food applications, pharmaceuticals, textiles, and biomedicine. Overexpression of soluble and active mTG in E. coli has been limited due to improper protein folding and requirement for proteolytic cleavage of the pro-domain. Furthermore, to integrate mTG more fully industrially and academically, thermostable and solvent-stable variants may be imperative...
February 28, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28241810/enhancing-the-thermostability-of-%C3%AE-l-rhamnosidase-from-aspergillus-terreus-and-the-enzymatic-conversion-of-rutin-to-isoquercitrin-by-adding-sorbitol
#18
Lin Ge, Anna Chen, Jianjun Pei, Linguo Zhao, Xianying Fang, Gang Ding, Zhenzhong Wang, Wei Xiao, Feng Tang
BACKGROUND: Thermally stable α-L-rhamnosidase with cleaving terminal α-L-rhamnose activity has great potential in industrial application. Therefore, it is necessary to find a proper method to improve the thermal stability of α-L-rhamnosidase. RESULTS: In this study, addition of sorbitol has been found to increase the thermostability of α-L-rhamnosidase from Aspergillus terreus at temperatures ranging from 65 °C to 80 °C. Half-life and activation free energy with addition of 2...
February 27, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28231788/economic-co-production-of-poly-malic-acid-and-pullulan-from-jerusalem-artichoke-tuber-by-aureobasidium-pullulans-ha-4d
#19
Jun Xia, Jiaxing Xu, Xiaoyan Liu, Jiming Xu, Xingfeng Wang, Xiangqian Li
BACKGROUND: poly(L-malic acid) (PMA) is a water-soluble polyester with many attractive properties in medicine and food industries, but the high cost of PMA fermentation has restricted its further application for large-scale production. To overcome this problem, PMA production from Jerusalem artichoke tubers was successfully performed. Additionally, a valuable exopolysaccharide, pullulan, was co-produced with PMA by Aureobasidum pullulans HA-4D. RESULTS: The Jerusalem artichoke medium for PMA and pullulan co-production contained only 100 g/L hydrolysate sugar, 30 g/L CaCO3 and 1 g/L NaNO3...
February 23, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28231778/expression-and-characteristics-of-manganese-peroxidase-from-ganoderma-lucidum-in-pichia-pastoris-and-its-application-in-the-degradation-of-four-dyes-and-phenol
#20
Hui Xu, Meng-Yuan Guo, Yan-Hua Gao, Xiao-Hui Bai, Xuan-Wei Zhou
BACKGROUND: Manganese peroxidase (MnP) of white rot basidiomycetes, an extracellular heme enzyme, is part of a peroxidase superfamily that is capable of degrading the different phenolic compounds. Ganoderma, a white rot basidiomycete widely distributed worldwide, could secrete lignin-modifying enzymes (LME), including laccase (Lac), lignin peroxidases (LiP) and MnP. RESULTS: After the selection of a G. lucidum strain from five Ganoderma strains, the 1092 bp full-length cDNA of the MnP gene, designated as G...
February 23, 2017: BMC Biotechnology
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