Journal of Microbiology

Journal Articles

Enhanced Poly-γ-Glutamic Acid Production by a Newly Isolated Bacillus halotolerans F29: Xiaorong Sun, Yaoyu Cai, Dexin Wang; J. Microbiol. 2024;62(8):695-707. Published online August 20, 2024; DOI: https://doi.org/10.1007/s12275-024-00153-w

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Poly-γ-glutamic acid (γ-PGA) is a promising biopolymer for various applications. In this study, we isolated a novel γ-PGA-producing strain, Bacillus halotolerans F29. The one-factor-at-a-time method was used to investigate the influence of carbon sources, nitrogen sources, and culture parameters on γ-PGA production. The optimal carbon and nitrogen sources were sucrose and (NH₄)₂SO₄, respectively. The optimal culture conditions for γ-PGA production were determined to be 37 °C and a pH of 5.5. Response surface methodology was used to determine the optimum medium components: 77.6 g/L sucrose, 43.0 g/L monosodium glutamate, and 2.2 g/L K₂HPO₄. The γ-PGA titer increased significantly from 8.5 ± 0.3 g/L to 20.7 ± 0.7 g/L when strain F29 was cultivated in the optimized medium. Furthermore, the γ-PGA titer reached 50.9 ± 1.5 g/L with a productivity of 1.33 g/L/h and a yield of 2.23 g of γ-PGA/g of L-glutamic acid with the optimized medium in fed-batch fermentation. The maximum γ-PGA titer reached 45.3 ± 1.1 g/L, with a productivity of 1.06 g/L/h when molasses was used as a carbon source. It should be noted that the γ-PGA yield in this study was the highest of all reported studies, indicating great potential for the industrial production of γ-PGA.

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Transcriptomics-guided rational engineering in Bacillus licheniformis for enhancing poly-γ-glutamic acid biosynthesis using untreated molasses
Rui Han, Qian Zhong, Yifan Yan, Juan Wang, Yifan Zhu, Sha Li, Peng Lei, Rui Wang, Yibin Qiu, Zhengshan Luo, Hong Xu
International Journal of Biological Macromolecules.2024; 282: 137514. CrossRef

Diversity and Dynamics of Marine Arenicolous Fungi in Three Seasides of the Korean Peninsula: Jun Won Lee , Chang Wan Seo , Wonjun Lee , Ji Seon Kim , Ki Hyeong Park , Yoonhee Cho , Young Woon Lim; J. Microbiol. 2023;61(1):63-82. Published online January 30, 2023; DOI: https://doi.org/10.1007/s12275-023-00011-1

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Abstract

Various arenicolous fungal species have been detected from the beach sand in the coastal area. However, little has been revealed regarding their distribution and dynamics. To investigate the overall diversity of marine arenicolous fungi (MAFs) in Korea and whether the composition of MAFs is affected by ocean currents, we isolated and analyzed the fungal community from the western, southern, and eastern seasides of the Korean Peninsula. In total, 603 strains were isolated and identified as 259 species based on appropriate molecular markers for each genus (ITS, BenA, CaM, tef1, and act). The composition of MAFs showed differences among the seasides. Our results indicate that many MAFs inhabit the beach sand on the Korean Peninsula, and the composition of MAFs is also affected by ocean currents flowing along each coast.

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The genus Peniophora (Russulales, Basidiomycota) from Patagonia revisited
Mario Rajchenberg, Andrés de Errasti, Sergio Pérez Gorjón
Mycological Progress.2024;[Epub] CrossRef
Contributions to the Inocybe umbratica–paludinella (Agaricales) Group in China: Taxonomy, Species Diversity, and Molecular Phylogeny
Xin Chen, Wen-Jie Yu, Tolgor Bau, P. Brandon Matheny, Egon Horak, Yu Liu, Li-Wu Qin, Li-Ping Tang, Yu-Peng Ge, Tie-Zhi Liu, Yu-Guang Fan
Journal of Fungi.2024; 10(12): 893. CrossRef
Long-Term Investigation of Marine-Derived Aspergillus Diversity in the Republic of Korea
Jun Won Lee, Wonjun Lee, Rekhani Hansika Perera, Young Woon Lim
Mycobiology.2023; 51(6): 436. CrossRef

Spot 42 RNA regulates putrescine catabolism in Escherichia coli by controlling the expression of puuE at the post-transcription level: Xin Sun , Ruyan Li , Guochen Wan , Wanli Peng , Shuangjun Lin , Zixin Deng , Rubing Liang; J. Microbiol. 2021;59(2):175-185. Published online February 1, 2021; DOI: https://doi.org/10.1007/s12275-021-0421-4

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Abstract

Putrescine, a typical polyamine compound important for cell growth and stress resistance, can be utilized as an energy source. However, the regulation of its catabolism is unclear. Here the small RNA (sRNA) Spot 42, an essential regulator of carbon catabolite repression (CCR), was confirmed to participate in the post-transcriptional regulation of putrescine catabolism in Escherichia coli. Its encoding gene spf exclusively exists in the γ-proteobacteria and contains specific binding sites to the 5􍿁-untranslated regions of the puuE gene, which encodes transaminase in the glutamylated putrescine pathway of putrescine catabolism converting γ-aminobutyrate (GABA) into succinate semialdehyde (SSA). The transcription of the spf gene was induced by glucose, inhibited by putrescine, and unaffected by PuuR, the repressor of puu genes. Excess Spot 42 repressed the expression of PuuE significantly in an antisense mechanism through the direct and specific base-pairing between the 51–57 nt of Spot 42 and the 5􍿁- UTR of puuE. Interestingly, Spot 42 mainly influenced the stability of the puuCBE transcript. This work revealed the regulatory role of Spot 42 in putrescine catabolism, in the switch between favorable and non-favorable carbon source utilization, and in the balance of metabolism of carbon and nitrogen sources.

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Regulation of TCA cycle genes by srbA sRNA: Impacts on Pseudomonas aeruginosa virulence and survival
Piyali Saha, Samir Kumar Mukherjee, Sk Tofajjen Hossain
Biochemical and Biophysical Research Communications.2024; 737: 150520. CrossRef
Rational Design of High-Efficiency Synthetic Small Regulatory RNAs and Their Application in Robust Genetic Circuit Performance Through Tight Control of Leaky Gene Expression
Jun Ren, Nuong Thi Nong, Phuong N. Lam Vo, Hyang-Mi Lee, Dokyun Na
ACS Synthetic Biology.2024; 13(10): 3256. CrossRef

The effects of deletion of cellobiohydrolase genes on carbon source-dependent growth and enzymatic lignocellulose hydrolysis in Trichoderma reesei: Meibin Ren , Yifan Wang , Guoxin Liu , Bin Zuo , Yuancheng Zhang , Yunhe Wang , Weifeng Liu , Xiangmei Liu , Yaohua Zhong; J. Microbiol. 2020;58(8):687-695. Published online June 10, 2020; DOI: https://doi.org/10.1007/s12275-020-9630-5

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Abstract

The saprophytic fungus Trichoderma reesei has long been used as a model to study microbial degradation of lignocellulosic biomass. The major cellulolytic enzymes of T. reesei are the cellobiohydrolases CBH1 and CBH2, which constitute more than 70% of total proteins secreted by the fungus. However, their physiological functions and effects on enzymatic hydrolysis of cellulose substrates are not sufficiently elucidated. Here, the cellobiohydrolase-encoding genes cbh1 and cbh2 were deleted, individually or combinatively, by using an auxotrophic marker-recycling technique in T. reesei. When cultured on media with different soluble carbon sources, all three deletion strains (Δcbh1, Δcbh2, and Δcbh1Δcbh2) exhibited no dramatic variation in morphological phenotypes, but their growth rates increased apparently when cultured on soluble cellulase-inducing carbon sources. In addition, Δcbh1 showed dramatically reduced growth and Δcbh1Δcbh2 could hardly grew on microcrystalline cellulose (MCC), whereas all strains grew equally on sodium carboxymethyl cellulose (CMC-Na), suggesting that the influence of the CBHs on growth was carbon source-dependent. Moreover, five representative cellulose substrates were used to analyse the influence of the absence of CBHs on saccharification efficiency. CBH1 deficiency significantly affected the enzymatic hydrolysis rates of various cellulose substrates, where acid pre-treated corn stover (PCS) was influenced the least. CBH2 deficiency reduced the hydrolysis of MCC, PCS, and acid pre-treated and delignified corncob but improved the hydrolysis ability of filter paper. These results demonstrate the specific contributions of CBHs to the hydrolysis of different types of biomass, which could facilitate the development of tailor-made strains with highly efficient hydrolysis enzymes for certain biomass types in the biofuel industry.

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An efficient CRISPR/Cas9 genome editing system based on a multiple sgRNA processing platform in Trichoderma reesei for strain improvement and enzyme production
Jiaxin Zhang, Kehang Li, Yu Sun, Cheng Yao, Weifeng Liu, Hong Liu, Yaohua Zhong
Biotechnology for Biofuels and Bioproducts.2024;[Epub] CrossRef
Transcriptome-wide analysis of a superior xylan degrading isolate Penicillium oxalicum 5–18 revealed active lignocellulosic degrading genes
Shuang Hu, Pei Han, Bao-Teng Wang, Long Jin, Hong-Hua Ruan, Feng-Jie Jin
Archives of Microbiology.2024;[Epub] CrossRef
Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass
Fernanda Lopes de Figueiredo, Fabiano Jares Contesini, César Rafael Fanchini Terrasan, Jaqueline Aline Gerhardt, Ana Beatriz Corrêa, Everton Paschoal Antoniel, Natália Sayuri Wassano, Lucas Levassor, Sarita Cândida Rabelo, Telma Teixeira Franco, Uffe Hasb
Microbial Cell Factories.2024;[Epub] CrossRef
Constitutive overexpression of cellobiohydrolase 2 in Trichoderma reesei reveals its ability to initiate cellulose degradation
Yubo Wang, Meibin Ren, Yifan Wang, Lu Wang, Hong Liu, Mei Shi, Yaohua Zhong
Engineering Microbiology.2023; 3(1): 100059. CrossRef
Inducer-free recombinant protein production in Trichoderma reesei: secretory production of endogenous enzymes and heterologous nanobodies using glucose as the sole carbon source
Toshiharu Arai, Mayumi Wada, Hiroki Nishiguchi, Yasushi Takimura, Jun Ishii
Microbial Cell Factories.2023;[Epub] CrossRef
The Influence of Trctf1 Gene Knockout by CRISPR–Cas9 on Cellulase Synthesis by Trichoderma reesei with Various Soluble Inducers
Yudian Chen, Yushan Gao, Zancheng Wang, Nian Peng, Xiaoqin Ran, Tingting Chen, Lulu Liu, Yonghao Li
Fermentation.2023; 9(8): 746. CrossRef
The effect of cellobiohydrolase 1 gene knockout for composition and hydrolytic activity of the enzyme complex secreted by filamentous fungus Penicillium verruculosum
Valeriy Yu. Kislitsin, Andrey M. Chulkin, Ivan N. Zorov, Yuri А. Denisenko, Arkadiy P. Sinitsyn, Alexandra M. Rozhkova
Bioresource Technology Reports.2022; 18: 101023. CrossRef
Deciphering the efficient cellulose degradation by the thermophilic fungus Myceliophthora thermophila focused on the synergistic action of glycoside hydrolases and lytic polysaccharide monooxygenases
Xing Qin, Jiahuan Zou, Kun Yang, Jinyang Li, Xiaolu Wang, Tao Tu, Yuan Wang, Bin Yao, Huoqing Huang, Huiying Luo
Bioresource Technology.2022; 364: 128027. CrossRef

Comparative portrayal of ocular surface microbe with and without dry eye: ZhenHao Li , Yufang Gong , ShuZe Chen , SiQi Li , Yu Zhang , HuiMin Zhong , ZhouCheng Wang , YiFan Chen , QiXin Deng , YuTing Jiang , LiYing Li , Min Fu , GuoGuo Yi; J. Microbiol. 2019;57(11):1025-1032. Published online August 28, 2019; DOI: https://doi.org/10.1007/s12275-019-9127-2

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Abstract

To compare the ocular surface (OS) microbial communities and diversity between dry eye (DE) and non-DE (NDE). Furthermore, we compared meibomian gland dysfunction (MGD) and non-MGD (NMGD) among DE subjects. The V3-V4 region of 16S rRNA gene high-throughput sequencing was performed in the conjunctival swab samples to investigate the composition of the OS bacterial community in DE (n=35) and NDE (n=54) and compared the composition of MGD (n=25) and NMGD (n=10) among DE subjects. Deep sequencing of OS 16S rDNA from DE (n=35) and NDE (n=54) demonstrated great a difference in alpha and beta diversity between the OS bacterial flora (P < 0.05). The similar OS microbial structures were shown at the phylum and genus levels by bioinformatics analysis between them, and in LEfSe (linear discriminant analysis effect size) analysis, Bacteroidia and Bacteroidetes were enriched in DE, while Pseudomonas was plentiful in NDE (linear discriminant analysis [LDA] > 4.0). Among the DE group, there was no significant difference in α and β diversity between MGD and NMGD (P > 0.05). Surprisingly, Bacilli was the dominant microbe in MGD, and Bacteroidetes was the superior bacteria in NMGD among DE subjects (LDA > 4.0). Different diversity of OS bacteria composition between DE and NDE and the altered diversity of OS bacteria may play an important role in DE. Moreover, the lower dominance of OS bacteria in DE may be associated with the occurrence and development of DE. Although there was no significant difference in alpha and beta analysis, the OS dominant microbe between MGD and NMGD among DE was different.

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Pasquale Aragona, Christophe Baudouin, Jose M. Benitez del Castillo, Elisabeth Messmer, Stefano Barabino, Jesus Merayo-Lloves, Francoise Brignole-Baudouin, Leandro Inferrera, Maurizio Rolando, Rita Mencucci, Maria Rescigno, Stefano Bonini, Marc Labetoulle
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Gramella fulva sp. nov., isolated from a dry surface of tidal flat: Sae Hyun Hwang , Woon Mo Hwang , Keunsoo Kang , Tae-Young Ahn; J. Microbiol. 2019;57(1):23-29. Published online November 19, 2018; DOI: https://doi.org/10.1007/s12275-019-8370-x

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Abstract

A novel Gram-stain-negative, aerobic, motile by means of gliding, and short rod-shaped bacterium, designated strain SH35T, was isolated from the dry surface of a tidal flat in Hwasung-si, South Korea. Growth occurred at 10–40°C (optimum 30°C), at pH 6.0–8.0 (optimum pH 7.0), in 1–12% NaCl (optimum 2%), and was inhibited in the absence of NaCl and Ca2+ ions. Phylogenetic analysis based on the 16S rRNA gene sequences showed that strain SH35T belonged to the genus Gramella and was a member of the family Flavobacteriaceae with highest sequence similarity to Gramella flava JLT2011T (96.1%), followed by Gramella oceani CCAMSZ- TT (95.6%), and 93.0–94.9% to other recognized Gramella species. The major cellular fatty acids (> 5% of the total) of strain SH35T were iso-C15:0, Iso-C16:0, anteiso-C15:0, iso-C17:0 3-OH and summed feature 9 (C16:0 10-methyl and/or C17:1 iso ω9с). The major polar lipids were phosphatidylethanolamine, two unidentified aminolipids and nine unidentified polar lipids. The major respiratory quinone and the predominant polyamine were menaquinone-6 (MK-6) and symhomospermidine, respectively. The DNA G + C content was 40.5 mol% (39.7% based on total genome calculations). Based on phylogenetic analysis and physiological and biochemical characterization, strain SH35T represents a novel species of the genus Gramella, for which the name Gramella fulva sp. nov. is proposed. The type strain is SH35T (= KACC 19447T = JCM 32369T).

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Optimization of Enterobacter cloacae (KU923381) for diesel oil degradation using Response Surface Methodology (RSM): Sugumar Ramasamy , Arumugam Arumugam , Preethy Chandran; J. Microbiol. 2017;55(2):104-111. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6265-2

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Abstract

Efficiency of Enterobacter cloacae KU923381 isolated from petroleum hydrocarbon contaminated soil was evaluated in batch culture and bioreactor mode. The isolate were screened for biofilm formation using qualitative and quantitative assays. Response surface methodology (RSM) was used to study the effect of pH, temperature, glucose concentration, and sodium chloride on diesel degradation. The predicted values for diesel oil degradation efficiency by the statistical designs are in a close agreement with experimental data (R2 = 99.66%). Degradation efficiency is increased by 36.78% at pH = 7, temperature = 35°C, glucose = 5%, and sodium chloride concentration = 5%. Under the optimized conditions, the experiments were performed for diesel oil degradation by gas chromatographic mass spectrometric analysis (GC-MS). GC-MS analysis confirmed that E. cloacae had highly degrade hexadecane, heptadecane, tridecane, and docosane by 99.71%, 99.23%, 99.66%, and 98.34% respectively. This study shows that rapid bioremoval of hydrocarbons in diesel oil is acheived by E. cloacae with abet of biofilm formation. The potential use of the biofilms for preparing trickling filters (gravel particles) for the degradation of hydrocarbons from petroleum wastes before their disposal in the open environment is highly suggested. This is the first successful attempt for artificially establishing petroleum hydrocarbon degrading bacterial biofilm on solid substrates in bioreactor.

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Review

MINIREVIEW] High-resolution imaging of the microbial cell surface: Ki Woo Kim; J. Microbiol. 2016;54(11):703-708. Published online October 29, 2016; DOI: https://doi.org/10.1007/s12275-016-6348-5

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Abstract

Microorganisms, or microbes, can function as threatening pathogens that cause disease in humans, animals, and plants; however, they also act as litter decomposers in natural ecosystems. As the outermost barrier and interface with the environment, the microbial cell surface is crucial for cell-to-cell communication and is a potential target of chemotherapeutic agents. Surface ultrastructures of microbial cells have typically been observed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Owing to its characteristics of low-temperature specimen preparation and superb resolution (down to 1 nm), cryo-field emission SEM has revealed paired rodlets, referred to as hydrophobins, on the cell walls of bacteria and fungi. Recent technological advances in AFM have enabled high-speed live cell imaging in liquid at the nanoscale level, leading to clear visualization of celldrug interactions. Platinum-carbon replicas from freeze-fractured fungal spores have been observed using transmission electron microscopy, revealing hydrophobins with varying dimensions. In addition, AFM has been used to resolve bacteriophages in their free state and during infection of bacterial cells. Various microscopy techniques with enhanced spatial resolution, imaging speed, and versatile specimen preparation are being used to document cellular structures and events, thus addressing unanswered biological questions.

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Journal Article

Application of Response Surface Methodology for Rapid Chrysene Biodegradation by Newly Isolated Marine-derived Fungus Cochliobolus lunatus Strain CHR4D: Jwalant K. Bhatt , Chirag M. Ghevariya , Dushyant R. Dudhagara , Rahul K. Rajpara , Bharti P. Dave; J. Microbiol. 2014;52(11):908-917. Published online October 31, 2014; DOI: https://doi.org/10.1007/s12275-014-4137-6

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Abstract

For the first time, Cochliobolus lunatus strain CHR4D, a marine-derived ascomycete fungus isolated from historically contaminated crude oil polluted shoreline of Alang-Sosiya ship-breaking yard, at Bhavnagar coast, Gujarat has been reported showing the rapid and enhanced biodegradation of chrysene, a four ringed high molecular weight (HMW) polycyclic aromatic hydrocarbon (PAH). Mineral Salt Broth (MSB) components such as ammonium tartrate and glucose along with chrysene, pH and trace metal solution have been successfully optimized by Response Surface Methodology (RSM) using central composite design (CCD). A validated, two-step optimization protocol has yielded a substantial 93.10% chrysene degradation on the 4th day, against unoptimized 56.37% degradation on the 14th day. The results depict 1.65 fold increase in chrysene degradation and 1.40 fold increase in biomass with a considerable decrement in time. Based on the successful laboratory experiments, C. lunatus strain CHR4D can thus be predicted as a potential candidate for mycoremediation of HMW PAHs impacted environments.

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Research Support, Non-U.S. Gov'ts

Surface Display Expression of Bacillus licheniformis Lipase in Escherichia coli Using Lpp’OmpA Chimera: Jae-Hyung Jo , Chan-Wook Han , Seung-Hwan Kim , Hyuk-Jin Kwon , Hyune-Hwan Lee; J. Microbiol. 2014;52(10):856-862. Published online August 27, 2014; DOI: https://doi.org/10.1007/s12275-014-4217-7

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Abstract

The lipase from Bacillus licheniformis ATCC14580 was displayed on the cell surface of Escherichia coli using Lpp’OmpA as the anchoring protein. The expressed Lpp’OmpA-lipase fusion protein has a molecular weight of approximately 35 kDa, which was confirmed by SDS-PAGE and western blot analysis. The Lpp’OmpA-lipase fusion protein was located on the cell surface, as determined by immunofluorescence confocal microscopy and flow cytometry. The enzyme activity of the surface-displayed lipase showed clear halo around the colony. The cell surface-displayed lipase showed the highest activity of 248.12 ± 9.42 U/g (lyophilized cell) at the optimal temperature of 37°C and pH 8.0. The enzyme exhibited the highest activity toward the substrate p-nitrophenyl caprylate (C8). These results suggest that E. coli, which displayed the lipase on its surface, could be used as a whole cell biocatalyst.

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Enhanced Production of Carboxymethylcellulase by a Marine Bacterium, Bacillus velezensis A-68, by Using Rice Hulls in Pilot-scale Bioreactor under Optimized Conditions for Dissolved Oxygen: Wa Gao , Hye-Jin Kim , Chung-Han Chung , Jin-Woo Lee; J. Microbiol. 2014;52(9):755-761. Published online July 30, 2014; DOI: https://doi.org/10.1007/s12275-014-4156-3

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Abstract

The optimal conditions for the production of carboxymethylcellulase (CMCase) by Bacillus velezensis A-68 at a flask scale have been previously reported. In this study, the parameters involved in dissolved oxygen in 7 and 100 L bioreactors were optimized for the pilot-scale production of CMCase. The optimal agitation speed and aeration rate for cell growth of B. velezensis A-68 were 323 rpm and 1.46 vvm in a 7 L bioreactor, whereas those for the production of CMCase were 380 rpm and 0.54 vvm, respectively. The analysis of variance (ANOVA) implied that the highly significant factor for cell growth was the aeration rate, whereas that for the production of CMCase was the agitation speed. The optimal inner pressures for cell growth and the production of CMCase by B. velezensis A-68 in a 100 L bioreactor were 0.00 and 0.04 MPa, respectively. The maximal production of CMCase in a 100 L bioreactor under optimized conditions using rice hulls was 108.1 U/ml, which was 1.8 times higher than that at a flask scale under previously optimized conditions.

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Journal Article

Application of Statistical Experimental Design for Optimization of Silver Nanoparticles Biosynthesis by a Nanofactory Streptomyces viridochromogenes: Noura El-Ahmady El-Naggar , Nayera A.M. Abdelwahed; J. Microbiol. 2014;52(1):53-63. Published online January 4, 2014; DOI: https://doi.org/10.1007/s12275-014-3410-z

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Abstract

Central composite design was chosen to determine the combined effects of four process variables (AgNO3 concentration, incubation period, pH level and inoculum size) on the extracellular biosynthesis of silver nanoparticles (AgNPs) by Streptomycesviridochromogenes. Statistical analysis of the results showed that incubation period, initial pH level and inoculum size had significant effects (P􌥑0.05) on the biosynthesis of silver nanoparticles at their individual level. The maximum biosynthesis of silver nanoparticles was achieved at a concentration of 0.5% (v/v) of 1 mM AgNO3, incubation period of 96 h, initial pH of 9 and inoculum size of 2% (v/v). After optimization, the biosynthesis of silver nanoparticles was improved by approximately 5-fold as compared to that of the unoptimized conditions. The synthetic process of silver nanoparticle generation using the reduction of aqueous Ag+ ion by the culture supernatants of S. viridochromogenes was quite fast, and silver nanoparticles were formed immediately by the addition of AgNO3 solution (1 mM) to the cell-free supernatant. Initial characterization of silver nanoparticles was performed by visual observation of color change from yellow to intense brown color. UV-visible spectrophotometry for measuring surface plasmon resonance showed a single absorption peak at 400 nm, which confirmed the presence of silver nanoparticles. Fourier Transform Infrared Spectroscopy analysis provided evidence for proteins as possible reducing and capping agents for stabilizing the nanoparticles. Transmission Electron Microscopy revealed the extracellular formation of spherical silver nanoparticles in the size range of 2.15–7.27 nm. Compared to the cell-free supernatant, the biosynthesized AgNPs revealed superior antimicrobial activity against Gram-negative, Gram-positive bacterial strains and Candida albicans.

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Surface Display of the HPV L1 Capsid Protein by the Autotransporter Shigella IcsA: Dan Xu , Xiaofeng Yang , Depu Wang , Jun Yu , Yili Wang; J. Microbiol. 2014;52(1):77-82. Published online January 4, 2014; DOI: https://doi.org/10.1007/s12275-014-3235-9

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Abstract

Autotransporters have become attractive tools for surface expression of foreign proteins in Gram-negative bacteria. In this study, the Shigella autotransporter IcsA, has been exploited to express the human papillomavirus (HPV) type 16 L1 capsid protein in Shigella sonnei and Escherichia coli. The L1 gene was fused in-frame to replace the coding sequence of the IcsA passenger domain that is responsible for actin-based motility. The resultant hybrid protein could be detected by an anti-L1 antibody on the surface of S. sonnei and E. coli. In E. coli, the protein was expressed on the entire surface of the bacterium. In contrast, the protein was detected mainly at one pole of the Shigella bacterium. However, the protein became evenly distributed on the surface of the Shigella bacterium when the icsP gene was removed. Our study demonstrated the possibility of exploiting autotransporters for surface expression of large, heterologous viral proteins, which may be a useful strategy for vaccine development.

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The Trimeric Autotransporter Adhesin SadA from Salmonella spp. as a Novel Bacterial Surface Display System
Shuli Sang, Wenge Song, Lu Lu, Qikun Ou, Yiyan Guan, Haoxia Tao, Yanchun Wang, Chunjie Liu
Vaccines.2024; 12(4): 399. CrossRef
The Shigella ProU system is required for osmotic tolerance and virulence
Rasha Y. Mahmoud, Wenqin Li, Ramadan A. Eldomany, Mohamed Emara, Jun Yu
Virulence.2017; 8(4): 362. CrossRef
The Multivalent Adhesion Molecule SSO1327 plays a key role in Shigella sonnei pathogenesis
Rasha Y. Mahmoud, Daniel Henry Stones, Wenqin Li, Mohamed Emara, Ramadan A. El‐domany, Depu Wang, Yili Wang, Anne Marie Krachler, Jun Yu
Molecular Microbiology.2016; 99(4): 658. CrossRef

Cell-Surface Expression of Aspergillus saitoi-Derived Functional α-1,2-Mannosidase on Yarrowia lipolytica for Glycan Remodeling: Hye Yun Moon , Trinh Luu Van , Seon Ah Cheon , Jinho Choo , Jeong-Yoon Kim , Hyun Ah Kang; J. Microbiol. 2013;51(4):506-514. Published online August 30, 2013; DOI: https://doi.org/10.1007/s12275-013-3344-x

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Abstract

Expression of proteins on the surface of yeast has a wide range of applications, such as development of live vaccines, screening of antibody libraries, and use as whole-cell biocatalysts. The hemiascomycetes yeast Yarrowia lipolytica has been raised as a potential host for heterologous expression of recombinant proteins. In this study, we report the expression of Aspergillus saitoi α-1,2-mannosidase, encoded by the msdS gene, on the cell surface of Y. lipolytica. As the first step to achieve the secretory expression of msdS protein, four different signal sequences-derived from the endogenous Y. lipolytica Lip2 and Xpr2 prepro regions and the heterologous A. niger α-amylase and rice α-amylase signal sequences-were analyzed for their secretion efficiency. It was shown that the YlLip2 prepro sequence was most efficient in directing the secretory expression of msdS in fully N-glycosylated forms. The surface display of msdS was subsequently directed by fusing GPI anchoring motifs derived from Y. lipolytica cell wall proteins, YlCwp1p and YlYwp1p, respectively, to the C-terminus of the Lip2 prepro-msdS protein. The expression of actively functional msdS protein on the cell surface was confirmed by western blot, flow cytometry analysis, along with the α-1,2-mannosidase activity assay using intact Y. lipolytica cells as the enzyme source. Furthermore, the glycoengineered Y. lipolytica Δoch1Δmpo1 strains displaying α-1,2-mannosidase were able to convert Man8GlcNAc2 to Man5GlcNAc2 efficiently on their cell-wall mannoproteins, demonstrating its potential used for glycoengineering in vitro or in vivo.

Citations

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Engineering novel Yarrowia lipolytica whole-cell biocatalysts by cell surface display of the native Lip2 lipase for biodiesel production
Maria Orfanidou, Eleftheria Panagiotidou, Antonios M. Makris, Eleni Theodosiou
Biotechnology for the Environment.2025;[Epub] CrossRef
Yeast Surface Display System: Strategies for Improvement and Biotechnological Applications
Karla V. Teymennet-Ramírez, Fernando Martínez-Morales, María R. Trejo-Hernández
Frontiers in Bioengineering and Biotechnology.2022;[Epub] CrossRef
Dietary intake of GDF11 delays the onset of several biomarkers of aging in male mice through anti-oxidant system via Smad2/3 pathway
Lili Song, Fei Wu, Congjun Li, Shicui Zhang
Biogerontology.2022; 23(3): 341. CrossRef
Hydrolytic secretome engineering in Yarrowia lipolytica for consolidated bioprocessing on polysaccharide resources: review on starch, cellulose, xylan, and inulin
Ewelina Celińska, Jean-Marc Nicaud, Wojciech Białas
Applied Microbiology and Biotechnology.2021; 105(3): 975. CrossRef
Yarrowia lipolytica Strains and Their Biotechnological Applications: How Natural Biodiversity and Metabolic Engineering Could Contribute to Cell Factories Improvement
Catherine Madzak
Journal of Fungi.2021; 7(7): 548. CrossRef
Engineering Yarrowia lipolytica for Use in Biotechnological Applications: A Review of Major Achievements and Recent Innovations
Catherine Madzak
Molecular Biotechnology.2018; 60(8): 621. CrossRef
Synthetic biology tools for engineering Yarrowia lipolytica
M. Larroude, T. Rossignol, J.-M. Nicaud, R. Ledesma-Amaro
Biotechnology Advances.2018; 36(8): 2150. CrossRef
Development of recombinant Yarrowia lipolytica producing virus-like particles of a fish nervous necrosis virus
Van-Trinh Luu, Hye Yun Moon, Jee Youn Hwang, Bo-Kyu Kang, Hyun Ah Kang
Journal of Microbiology.2017; 55(8): 655. CrossRef
Using a vector pool containing variable-strength promoters to optimize protein production in Yarrowia lipolytica
Rémi Dulermo, François Brunel, Thierry Dulermo, Rodrigo Ledesma-Amaro, Jérémy Vion, Marion Trassaert, Stéphane Thomas, Jean-Marc Nicaud, Christophe Leplat
Microbial Cell Factories.2017;[Epub] CrossRef
Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering
Catherine Madzak
Applied Microbiology and Biotechnology.2015; 99(11): 4559. CrossRef
Biotechnological applications of Yarrowia lipolytica: Past, present and future
Hu-Hu Liu, Xiao-Jun Ji, He Huang
Biotechnology Advances.2015; 33(8): 1522. CrossRef
Functional characterization of extracellular chitinase encoded by the YlCTS1 gene in a dimorphic yeast Yarrowia lipolytica
Jeong-Nam Park, Chang Pyo Han, Dong-Jik Lee, Seon Ah Cheon, Hyun Ah Kang
Journal of Microbiology.2014; 52(4): 284. CrossRef
Characterization of putative glycosylphosphatidylinositol-anchoring motifs for surface display in the methylotrophic yeast Hansenula polymorpha
Seon Ah Cheon, Jinhee Jung, Jin Ho Choo, Doo-Byoung Oh, Hyun Ah Kang
Biotechnology Letters.2014; 36(10): 2085. CrossRef

Research Support, U.S. Gov't, Non-P.H.S.

NOTE] Fosmid Cloning, Nucleotide Sequence, and Characterization of a Beta-Lactamase Gene from Subsurface Isolates: Nurcan Vardar , Gönül Vardar-Schara; J. Microbiol. 2012;50(4):680-683. Published online July 21, 2012; DOI: https://doi.org/10.1007/s12275-012-2139-9

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Abstract: A beta-lactamase gene was isolated for the first time from a terrestrial subsurface environment using a combined cultivation and direct cloning strategy. The gene, discovered from 24 m below land surface in Hawaii, was most similar to the penicillinase from Bacillus licheniformis. The resistance gene was confirmed via subcloning and its minimum inhibitory concentration values were measured against several test betalactam antibiotics. This study extends the knowledge on resistance to antimicrobials, which may help the efforts to minimize their future threat.

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