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Microbial Cell Factories

Sha Tao, Ying Qian, Xin Wang, Weijia Cao, Weichao Ma, Kequan Chen, Pingkai Ouyang
BACKGROUND: Microbial biosynthesis of natural products holds promise for preclinical studies and treating diseases. For instance, pinocembrin is a natural flavonoid with important pharmacologic characteristics and is widely used in preclinical studies. However, high yield of natural products production is often limited by the intracellular cofactor level, including adenosine triphosphate (ATP). To address this challenge, tailored modification of ATP concentration in Escherichia coli was applied in efficient pinocembrin production...
September 18, 2018: Microbial Cell Factories
Jyotiranjan Bal, Hee-Young Jung, Luong Ngoc Nguyen, Jisang Park, Yong-Suk Jang, Dae-Hyuk Kim
BACKGROUND: Dengue is a rapidly spreading mosquito borne tropical viral disease affecting hundreds of millions of people across the globe annually. The dengue virus (DENV) includes four genetically distinct serotypes that cause serious life-threatening infections, including dengue hemorrhagic fever/dengue shock syndrome. Dengue vaccine development is complicated by the possibility of vaccine-enhanced severe dengue disease due to antibody-dependent enhancement by pre-existing cross-reactivity, as well as homotypic antibodies...
September 14, 2018: Microbial Cell Factories
Jian Zha, Ying Zang, Matthew Mattozzi, Jens Plassmeier, Mamta Gupta, Xia Wu, Sonya Clarkson, Mattheos A G Koffas
BACKGROUND: Anthocyanins such as cyanidin 3-O-glucoside (C3G) have wide applications in industry as food colorants. Their current production heavily relies on extraction from plant tissues. Development of a sustainable method to produce anthocyanins is of considerable interest for industrial use. Previously, E. coli-based microbial production of anthocyanins has been investigated extensively. However, safety concerns on E. coli call for the adoption of a safe production host. In the present study, a GRAS bacterium, Corynebacterium glutamicum, was introduced as the host strain to synthesize C3G...
September 14, 2018: Microbial Cell Factories
Yang Liu, Wen-Juan Mo, Tian-Fang Shi, Meng-Zhu Wang, Jun-Gang Zhou, Yao Yu, Wen-Shan Yew, Hong Lu
BACKGROUND: The yeast Kluyveromyces marxianus is an emerging cell factory for heterologous protein biosynthesis and its use holds tremendous advantages for multiple applications. However, which genes influence the productivity of desired proteins in K. marxianus has so far been investigated by very few studies. RESULTS: In this study, we constructed a K. marxianus recombinant (FIM1/Est1E), which expressed the heterologous ruminal feruloyl esterase Est1E as reporter...
September 14, 2018: Microbial Cell Factories
Coraline Rigouin, Christian Croux, Vinciane Borsenberger, Maher Ben Khaled, Thierry Chardot, Alain Marty, Florence Bordes
BACKGROUND: Oleaginous yeast Yarrowia lipolytica is an organism of choice for the development of biofuel and oleochemicals. It has become a chassis for metabolic engineering in order to produce targeted lipids. Understanding the function of key-enzymes involved in lipid metabolism is essential to design better routes for enhanced lipid production and for strains producing lipids of interest. Because medium chain fatty acids (MCFA) are valuable compounds for biokerosene production, we previously generated strains capable of producing MCFA up to 12% of total lipid content (Rigouin et al...
September 10, 2018: Microbial Cell Factories
Shuhong Mao, Jian-Wen Wang, Fufeng Liu, Zhangliang Zhu, Dengke Gao, Qianqian Guo, Panpan Xu, Zheng Ma, Yali Hou, Xiaotao Cheng, Dengyue Sun, Fuping Lu, Hui-Min Qin
BACKGROUND: Biosynthesis of steroidal drugs is of great benefit in pharmaceutical manufacturing as the process involves efficient enzymatic catalysis at ambient temperature and atmospheric pressure compared to chemical synthesis. 3-ketosteroid-∆1 -dehydrogenase from Arthrobacter simplex (KsdD3) catalyzes 1,2-desaturation of steroidal substrates with FAD as a cofactor. RESULTS: Recombinant KsdD3 exhibited organic solvent tolerance. W117, F296, W299, et al., which were located in substrate-binding cavity, were predicted to form hydrophobic interaction with the substrate...
September 10, 2018: Microbial Cell Factories
Matthew Hilliard, Andrew Damiani, Q Peter He, Thomas Jeffries, Jin Wang
BACKGROUND: Scheffersomyces stipitis is an important yeast species in the field of biorenewables due to its desired capacity for xylose utilization. It has been recognized that redox balance plays a critical role in S. stipitis due to the different cofactor preferences in xylose assimilation pathway. However, there has not been any systems level understanding on how the shift in redox balance contributes to the overall metabolic shift in S. stipitis to cope with reduced oxygen uptake...
September 5, 2018: Microbial Cell Factories
Laura Ortiz-Velez, Javiera Ortiz-Villalobos, Abby Schulman, Jee-Hwan Oh, Jan-Peter van Pijkeren, Robert A Britton
BACKGROUND: Lactic acid bacteria (LAB) are one of the microorganisms of choice for the development of protein delivery systems for therapeutic purposes. Although there are numerous tools to facilitate genome engineering of lactobacilli; transformation efficiency still limits the ability to engineer their genomes. While genetically manipulating Lactobacillus reuteri ATCC PTA 6475 (LR 6475), we noticed that after an initial transformation, several LR 6475 strains significantly improved their ability to take up plasmid DNA via electroporation...
September 3, 2018: Microbial Cell Factories
María-Efigenia Álvarez-Cao, Agustín Rico-Díaz, María-Esperanza Cerdán, Manuel Becerra, María-Isabel González-Siso
BACKGROUND: The recycling of agro-industrial wastes is at present limited by the availability of efficient and low-cost enzyme cocktails. The use of these materials as culture media to produce the enzymes can contribute to the profitability of the recycling process and to the circular economy. The aim of this work is the construction of a recombinant yeast strain efficient to grow in mixed whey (residue of cheese making) and beet molasses (residue of sugar manufacture) as culture medium, and to produce heterologous α-galactosidase, an enzyme with varied industrial applications and wide market...
September 3, 2018: Microbial Cell Factories
Romana Koszagova, Tomas Krajcovic, Klaudia Palencarova-Talafova, Vladimir Patoprsty, Alica Vikartovska, Kristyna Pospiskova, Ivo Safarik, Jozef Nahalka
BACKGROUND: Physiological aggregation of a recombinant enzyme into enzymatically active inclusion bodies could be an excellent strategy to obtain immobilized enzymes for industrial biotransformation processes. However, it is not convenient to recycle "gelatinous masses" of protein inclusion bodies from one reaction cycle to another, as high centrifugation forces are needed in large volumes. The magnetization of inclusion bodies is a smart solution for large-scale applications, enabling an easier separation process using a magnetic field...
September 3, 2018: Microbial Cell Factories
Jeffrey J Czajka, Justin A Nathenson, Veronica T Benites, Edward E K Baidoo, Qianshun Cheng, Yechun Wang, Yinjie J Tang
BACKGROUND: β-Ionone is a fragrant terpenoid that generates a pleasant floral scent and is used in diverse applications as a cosmetic and flavoring ingredient. A growing consumer desire for natural products has increased the market demand for natural β-ionone. To date, chemical extraction from plants remains the main approach for commercial natural β-ionone production. Unfortunately, changing climate and geopolitical issues can cause instability in the β-ionone supply chain. Microbial fermentation using generally recognized as safe (GRAS) yeast offers an alternative method for producing natural β-ionone...
September 1, 2018: Microbial Cell Factories
Kevin James Metcalf, Marilyn F Slininger Lee, Christopher Matthew Jakobson, Danielle Tullman-Ercek
Biotechnological processes use microbes to convert abundant molecules, such as glucose, into high-value products, such as pharmaceuticals, commodity and fine chemicals, and energy. However, from the outset of the development of a new bioprocess, it is difficult to determine the feasibility, expected yields, and targets for engineering. In this review, we describe a methodology that uses rough estimates to assess the feasibility of a process, approximate the expected product titer of a biological system, and identify variables to manipulate in order to achieve the desired performance...
August 30, 2018: Microbial Cell Factories
Haojie Cao, Oscar P Kuipers
BACKGROUND: Gram-positive bacterium Bacillus subtilis has been extensively studied as a microbial cell factory for high-level producing a wide range of interesting products. Green fluorescent protein (GFP) is commonly used as a marker for determining the strength of a given promoter or for the subcellular localization of a fusion protein. However, the inherent heterogeneity of GFP expression among individual cells that can arise from global regulation differences in the expression host, has not yet been systematically assessed...
August 30, 2018: Microbial Cell Factories
Huiling Cheng, Siqi Wang, Muhammad Bilal, Xuemei Ge, Can Zhang, Patrick Fickers, Hairong Cheng
BACKGROUND: Erythritol is a four-carbon sugar alcohol with sweetening properties that is used by the agro-food industry as a food additive. In the yeast Yarrowia lipolytica, the last step of erythritol synthesis involves the reduction of erythrose by specific erythrose reductase(s). In the earlier report, an erythrose reductase gene (YALI0F18590g) from erythritol-producing yeast Y. lipolytica MK1 was identified (Janek et al. in Microb Cell Fact 16:118, 2017). However, deletion of the gene in Y...
August 29, 2018: Microbial Cell Factories
L Kyselova, D Kreitmayer, A Kremling, K Bettenbrock
BACKGROUND: Glucose is the main carbon source of E. coli and a typical substrate in production processes. The main glucose uptake system is the glucose specific phosphotransferase system (Glc-PTS). The PTS couples glucose uptake with its phosphorylation. This is achieved by the concomitant conversion of phosphoenolpyruvate (PEP) to pyruvate. The Glc-PTS is hence unfavorable for the production of succinate as this product is derived from PEP. RESULTS: We studied, in a systematic manner, the effect of knocking out the Glc-PTS and of replacing it with the glucose facilitator (Glf) of Zymomonas mobilis on succinate yield and productivity...
August 28, 2018: Microbial Cell Factories
Yan Liu, Rong Gong, Xiaoqin Liu, Peichao Zhang, Qi Zhang, You-Sheng Cai, Zixin Deng, Margit Winkler, Jianguo Wu, Wenqing Chen
BACKGROUND: Tubercidin (TBN), an adenosine analog with potent antimycobacteria and antitumor bioactivities, highlights an intriguing structure, in which a 7-deazapurine core is linked to the ribose moiety by an N-glycosidic bond. However, the molecular logic underlying the biosynthesis of this antibiotic has remained poorly understood. RESULTS: Here, we report the discovery and characterization of the TBN biosynthetic pathway from Streptomyces tubercidicus NBRC 13090 via reconstitution of its production in a heterologous host...
August 28, 2018: Microbial Cell Factories
Xuan Xin, Mengmeng Zhang, Xiaofeng Li, Furao Lai, Guanglei Zhao
BACKGROUND: Flavonoid glycosides have many beneficial effects on health, but these bioactivities tend to decrease after oral administration owing to their poor lipophilicity. In this study, a facile whole-cell-based method was developed for selective preparation of monoester or diester of troxerutin, a flavonoid derivative. In addition, the bioavailabilities and antioxidant properties of troxerutin and its acylated derivatives were also investigated in cells. RESULTS: Pseudomonas aeruginosa and Pseudomonas stutzeri cells showed high catalytic efficiency (substrate conversion > 90%) and different preferences for troxerutin, resulting in the production of its monoester (TME) and diester (TDE), respectively...
August 22, 2018: Microbial Cell Factories
Kei-Anne Baritugo, Hee Taek Kim, Yokimiko David, Tae Uk Khang, Sung Min Hyun, Kyoung Hee Kang, Ju Hyun Yu, Jong Hyun Choi, Jae Jun Song, Jeong Chan Joo, Si Jae Park
BACKGROUND: Recent interest has been focused on the production of platform chemicals from renewable biomass due to increasing concerns on global warming and depletion of fossil fuel reserves. Microbial production of platform chemicals in biorefineries has been suggested to be a promising solution for these problems. Gamma-aminobutyrate (GABA), a versatile bulk chemical used in food and pharmaceutical industry, is also used as a key monomer for nylon 4. GABA can be biologically produced by decarboxylation of glutamate...
August 21, 2018: Microbial Cell Factories
Tutku Kurt, Anna-Maria Marbà-Ardébol, Zeynep Turan, Peter Neubauer, Stefan Junne, Vera Meyer
BACKGROUND: Filamentous fungi including Aspergillus niger are cell factories for the production of organic acids, proteins and bioactive compounds. Traditionally, stirred-tank reactors (STRs) are used to cultivate them under highly reproducible conditions ensuring optimum oxygen uptake and high growth rates. However, agitation via mechanical stirring causes high shear forces, thus affecting fungal physiology and macromorphologies. Two-dimensional rocking-motion wave-mixed bioreactor cultivations could offer a viable alternative to fungal cultivations in STRs, as comparable gas mass transfer is generally achievable while deploying lower friction and shear forces...
August 21, 2018: Microbial Cell Factories
Megumi Miyano, Kosei Tanaka, Shu Ishikawa, Kotaro Mori, Andrés Miguel-Arribas, Wilfried J J Meijer, Ken-Ichi Yoshida
BACKGROUND: Bacterial strains of the genus Geobacillus grow at high temperatures of 50-75 °C and could thus be useful for biotechnological applications. However, genetic manipulation of these species is difficult because the current techniques for transforming Geobacillus species are not efficient. In this study, we developed an easy and efficient method for transforming Geobacillus kaustophilus using the conjugative plasmid pLS20cat. RESULTS: We constructed a transformation system comprising (i) a mobilizable Bacillus subtilis-G...
August 17, 2018: Microbial Cell Factories
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