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Microbial Cell Factories

Wah Seng See-Too, Peter Convey, David A Pearce, Kok-Gan Chan
BACKGROUND: N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum sensing. We have previously demonstrated the degradation of AHLs by an Antarctic bacterium, Planococcus versutus L10.15T , at low temperature through the production of an AHL lactonase. In this study, we cloned the AHL lactonase gene and characterized the purified novel enzyme. RESULTS: Rapid resolution liquid chromatography analysis indicated that purified AidP possesses high AHL-degrading activity on unsubstituted, and 3-oxo substituted homoserine lactones...
November 17, 2018: Microbial Cell Factories
Katarzyna Ciaćma, Jerzy Więckiewicz, Sylwia Kędracka-Krok, Magdalena Kurtyka, Małgorzata Stec, Maciej Siedlar, Jarek Baran
BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively eliminates tumor cells. However, the short biological half-life of this molecule limits its potential use in the clinic. Our aim was to construct a recombinant strain of nonpathogenic Lactococcus lactis bacteria as a vector for effective and prolonged human TRAIL production. Herein, we examined the expression and secretion conditions leading to the production of biologically active protein in vitro...
November 16, 2018: Microbial Cell Factories
Cristiana Faria, Nuno Borges, Isabel Rocha, Helena Santos
BACKGROUND: Mannosylglycerate (MG) is one of the most widespread compatible solutes among marine microorganisms adapted to hot environments. This ionic solute holds excellent ability to protect proteins against thermal denaturation, hence a large number of biotechnological and clinical applications have been put forward. However, the current prohibitive production costs impose severe constraints towards large-scale applications. All known microbial producers synthesize MG from GDP-mannose and 3-phosphoglycerate via a two-step pathway in which mannosyl-3-phosphoglycerate is the intermediate metabolite...
November 16, 2018: Microbial Cell Factories
Honghua Li, Pengjie Hu, Ying Wang, Yuanyuan Pan, Gang Liu
BACKGROUND: Autophagy is used for degradation of cellular components and nutrient recycling. Atg8 is one of the core proteins in autophagy and used as a marker for autophagic detection. However, the autophagy of filamentous fungi is poorly understood compared with that of Saccharomyces cerevisiae. Our previous study revealed that disruption of the autophagy related gene Acatg1 significantly enhanced cephalosporin C yield through reducing degradation of cephalosporin biosynthetic proteins in Acremonium chrysogenum, suggesting that modulation of autophagic process is one promising way to increase antibiotic production in A...
November 13, 2018: Microbial Cell Factories
Seiya Takayama, Aiko Ozaki, Rie Konishi, Chisako Otomo, Mayumi Kishida, Yuuki Hirata, Takuya Matsumoto, Tsutomu Tanaka, Akihiko Kondo
BACKGROUND: Economical production of value-added chemicals from renewable biomass is a promising path to sustainability. 3-Hydroxypropionic acid (3-HP) is an important chemical for building a bio-sustainable society. Establishment of 3-HP production from renewable resources such as glucose would provide a bio-sustainable alternative to the production of acrylic acid from fossil resources. RESULTS: Here, we describe metabolic engineering of the fission yeast Schizosaccharomyces pombe to enhance 3-HP production from glucose and cellobiose via the malonyl-CoA pathway...
November 13, 2018: Microbial Cell Factories
Manuel Benedetti, Valeria Vecchi, Simone Barera, Luca Dall'Osto
Interest in bulk biomass from microalgae, for the extraction of high-value nutraceuticals, bio-products, animal feed and as a source of renewable fuels, is high. Advantages of microalgal vs. plant biomass production include higher yield, use of non-arable land, recovery of nutrients from wastewater, efficient carbon capture and faster development of new domesticated strains. Moreover, adaptation to a wide range of environmental conditions evolved a great genetic diversity within this polyphyletic group, making microalgae a rich source of interesting and useful metabolites...
November 10, 2018: Microbial Cell Factories
Songwei Wang, Cong Fu, Muhammad Bilal, Hongbo Hu, Wei Wang, Xuehong Zhang
BACKGROUND: Arbutin is a plant-derived glycoside with potential antioxidant, antibacterial and anti-inflammatory activities. Currently, it is mainly produced by plant extraction or enzymatic processes, which suffers from expensive processing cost and low product yield. Metabolic engineering of microbes is an increasingly powerful method for the high-level production of valuable biologicals. Since Pseudomonas chlororaphis has been widely engineered as a phenazine-producing platform organism due to its well-characterized genetics and physiology, and faster growth rate using glycerol as a renewable carbon source, it can also be engineered as the cell factory using strong shikimate pathway on the basis of synthetic biology...
November 10, 2018: Microbial Cell Factories
Yide Huang, Yanyu Long, Suhuan Li, Ting Lin, Jingwen Wu, Yafei Zhang, Yao Lin
BACKGROUND: Proprotein convertase furin is responsible for the processing of a wide variety of precursors consisted of signal peptide, propeptide and mature peptide in mammal. Many precursors processed by furin have important physiological functions and can be recombinantly expressed in Pichia pastoris expression system for research, pharmaceutical and vaccine applications. However, it is not clear whether the furin cleavage sites between the propeptide and mature peptide can be properly processed in P...
November 8, 2018: Microbial Cell Factories
Olga Marchut-Mikolajczyk, Piotr Drożdżyński, Dominika Pietrzyk, Tadeusz Antczak
BACKGROUND: The process of plant growth in the contaminated environment is often inhibited and entails the neutralization of harmful compounds. To reduce the negative impact of harmful compounds microorganisms produce unique compounds called biosurfactants. This paper describes the potential of culturable endophytic microorganisms from synanthropic plant-Chelidonium majus L. for the production of biosurfactants, as indirect plant promoting factors as well as their degradation activity...
November 3, 2018: Microbial Cell Factories
P Mara, G S Fragiadakis, F Gkountromichos, D Alexandraki
Ammonium assimilation is linked to fundamental cellular processes that include the synthesis of non-essential amino acids like glutamate and glutamine. In Saccharomyces cerevisiae glutamate can be synthesized from α-ketoglutarate and ammonium through the action of NADP-dependent glutamate dehydrogenases Gdh1 and Gdh3. Gdh1 and Gdh3 are evolutionarily adapted isoforms and cover the anabolic role of the GDH-pathway. Here, we review the role and function of the GDH pathway in glutamate metabolism and we discuss the additional contributions of the pathway in chromatin regulation, nitrogen catabolite repression, ROS-mediated apoptosis, iron deficiency and sphingolipid-dependent actin cytoskeleton modulation in S...
November 1, 2018: Microbial Cell Factories
Johanna Hausjell, Julia Weissensteiner, Christian Molitor, Heidi Halbwirth, Oliver Spadiut
BACKGROUND: Escherichia coli is one of the most widely used hosts for recombinant protein production in academia and industry. Strain BL21(DE3) is frequently employed due to its advantageous feature of lacking proteases which avoids degradation of target protein. Usually it is used in combination with the T7-pET system where induction is performed by one point addition of IPTG. We recently published a few studies regarding lactose induction in BL21(DE3) strains. BL21(DE3) can only take up the glucose-part of the disaccharide when fed with lactose...
October 30, 2018: Microbial Cell Factories
Phillipp Mueller, Rahul Gauttam, Nadja Raab, René Handrick, Claudia Wahl, Sebastian Leptihn, Michael Zorn, Michaela Kussmaul, Marianne Scheffold, Bernhard Eikmanns, Lothar Elling, Sabine Gaisser
BACKGROUND: Increasing efforts have been made to assess the potential of Escherichia coli strains for the production of complex recombinant proteins. Since a considerable part of therapeutic proteins are glycoproteins, the lack of the post-translational attachment of sugar moieties in standard E. coli expression strains represents a major caveat, thus limiting the use of E. coli based cell factories. The establishment of an E. coli expression system capable of protein glycosylation could potentially facilitate the production of therapeutics with a putative concomitant reduction of production costs...
October 26, 2018: Microbial Cell Factories
Lin Liu, Haoran Yu, Kun Du, Zhiyan Wang, Yiru Gan, He Huang
BACKGROUND: High thermostability is required for trypsin to have wider industrial applications. Target mutagenesis at flexible regions has been proved to be an efficient protein engineering method to enhance the protein thermostability. RESULTS: The flexible regions in porcine trypsin were predicted using the methods including molecular dynamic simulation, FlexPred, and FoldUnfold. The amino acids 78-90 was predicted to be the highly flexible region simultaneously by the three methods and hence selected to be the mutation target...
October 25, 2018: Microbial Cell Factories
Aiqun Yu, Yakun Zhao, Yaru Pang, Zhihui Hu, Cuiying Zhang, Dongguang Xiao, Matthew Wook Chang, Susanna Su Jan Leong
BACKGROUND: Microbial biofuel production provides a promising sustainable alternative to fossil fuels. 1-Butanol is recognized as an advanced biofuel and is gaining attention as an ideal green replacement for gasoline. In this proof-of-principle study, the oleaginous yeast Yarrowia lipolytica was first engineered with a heterologous CoA-dependent pathway and an endogenous pathway, respectively. RESULTS: The co-overexpression of two heterologous genes ETR1 and EutE resulted in the production of 1-butanol at a concentration of 65 μg/L...
October 25, 2018: Microbial Cell Factories
Lokanand Koduru, Meiyappan Lakshmanan, Dong-Yup Lee
BACKGROUND: Cellular metabolism is tightly regulated by hard-wired multiple layers of biological processes to achieve robust and homeostatic states given the limited resources. As a result, even the most intuitive enzyme-centric metabolic engineering endeavours through the up-/down-regulation of multiple genes in biochemical pathways often deliver insignificant improvements in the product yield. In this regard, targeted engineering of transcriptional regulators (TRs) that control several metabolic functions in modular patterns is an interesting strategy...
October 25, 2018: Microbial Cell Factories
Wolfgang Mischko, Max Hirte, Monika Fuchs, Norbert Mehlmer, Thomas B Brück
BACKGROUND: Terpenes are an important and extremely versatile class of secondary metabolites that are commercially used in the pharmaceutical, food and cosmetics sectors. Genome mining of different fungal collections has revealed the genetic basis for a steadily increasing number of putative terpene synthases without any detailed knowledge about their biochemical properties. The analysis and research of this rich genetic source provides a precious basis for the advancing biotechnological production of an almost endless number of valuable natural metabolites...
October 22, 2018: Microbial Cell Factories
Xin Liu, Hai Wang, Bin Wang, Li Pan
BACKGROUND: Bacillus subtilis has been widely used as a host for heterologous protein expression in food industry. B. subtilis ATCC6051 is an alternative expression host for the production of industrial enzymes, and exhibits favorable growth properties compared to B. subtilis 168. Extracellular expression of pullulanase from recombinant B. subtilis is still limited due to the issues on promoters of B. subtilis expression system. This study was undertaken to develop a new, high-level expression system in B...
October 22, 2018: Microbial Cell Factories
Susanne Katharina Schwechheimer, Judith Becker, Lindsay Peyriga, Jean-Charles Portais, Christoph Wittmann
BACKGROUND: The fungus Ashbya gossypii is an important industrial producer of the vitamin riboflavin. Using this microbe, riboflavin is manufactured in a two-stage process based on a rich medium with vegetable oil, yeast extract and different precursors: an initial growth and a subsequent riboflavin production phase. So far, our knowledge on the intracellular metabolic fluxes of the fungus in this complex process is limited, but appears highly relevant to better understand and rationally engineer the underlying metabolism...
October 16, 2018: Microbial Cell Factories
Juan J Barrero, Jason C Casler, Francisco Valero, Pau Ferrer, Benjamin S Glick
BACKGROUND: Proteins can be secreted from a host organism with the aid of N-terminal secretion signals. The budding yeast Pichia pastoris (Komagataella sp.) is widely employed to secrete proteins of academic and industrial interest. For this yeast, the most commonly used secretion signal is the N-terminal portion of pre-pro-α-factor from Saccharomyces cerevisiae. However, this secretion signal promotes posttranslational translocation into the endoplasmic reticulum (ER), so proteins that can fold in the cytosol may be inefficiently translocated and thus poorly secreted...
October 12, 2018: Microbial Cell Factories
Yang Hu, Yanjie Liu, Xiaoran Hao, Dan Wang, Oren Akhberdi, Biyun Xiang, Xudong Zhu
BACKGROUND: The canonical heterotrimeric G protein-cAMP/PKA pathway regulates numerous cellular processes in filamentous fungi. Chaetomium globosum, a saprophytic fungus, is known for producing many secondary metabolites, including cytotoxic chaetoglobosin A (ChA), as well as abundant cellulase and xylanase. RESULTS: Here we report on the functional characterization of this signaling pathway in C. globosum. We blocked the pathway by knocking down the putative Gα-encoding gene gna1 (in the pG14 mutant)...
October 11, 2018: Microbial Cell Factories
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