Journal of Microbiology

Minireview

A review on computational models for predicting protein solubility: Teerapat Pimtawong, Jun Ren, Jingyu Lee, Hyang-Mi Lee, Dokyun Na; J. Microbiol. 2025;63(1):e.2408001. Published online January 24, 2025; DOI: https://doi.org/10.71150/jm.2408001

868 View
116 Download

Abstract PDF: Protein solubility is a critical factor in the production of recombinant proteins, which are widely used in various industries, including pharmaceuticals, diagnostics, and biotechnology. Predicting protein solubility remains a challenging task due to the complexity of protein structures and the multitude of factors influencing solubility. Recent advances in computational methods, particularly those based on machine learning, have provided powerful tools for predicting protein solubility, thereby reducing the need for extensive experimental trials. This review provides an overview of current computational approaches to predict protein solubility. We discuss the datasets, features, and algorithms employed in these models. The review aims to bridge the gap between computational predictions and experimental validations, fostering the development of more accurate and reliable solubility prediction models that can significantly enhance recombinant protein production.

Journal Article

Comparative Secretory Efficiency of Two Chitosanase Signal Peptides from Bacillus subtilis in Escherichia coli: Tae-Yang Eom, Yehui Gang, Youngdeuk Lee, Yoon-Hyeok Kang, Eunyoung Jo, Svini Dileepa Marasinghe, Heung Sik Park, Gun-Hoo Park, Chulhong Oh; J. Microbiol. 2024;62(12):1155-1164. Published online November 25, 2024; DOI: https://doi.org/10.1007/s12275-024-00186-1

52 View
0 Download

Abstract: The production of recombinant proteins in Escherichia coli is often challenged by cytoplasmic expression due to proteolytic degradation and inclusion body formation. Extracellular expression can overcome these problems by simplifying downstream processing and improving protein yields. This study aims to compare the efficiency of two Bacillus subtilis chitosanase signal peptides in mediating extracellular secretion in E. coli. We identified a naturally occurring mutant signal peptide (mCsn2-SP) from B. subtilis CH2 chitosanase (CH2CSN), which is characterized by a deletion of six amino acids in the N-region relative to the signal peptide (Csn1-SP) from B. subtilis CH1 chitosanase (CH1CSN). The CH1CSN and CH2CSN genes were cloned into the pET-11a vector and protein secretion was evaluated in E. coli BL21(DE3) host cells. Expression was induced with 0.1 mM and 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 30 °C for one and three days. CH2CSN showed higher secretion levels compared to CH1CSN under all experimental conditions, especially with 0.1 mM IPTG induction for 3 days, which resulted in a 2.37-fold increase in secretion. Furthermore, it was demonstrated that mCsn2-SP is capable of secreting human Cu,Zn-superoxide dismutase (hSOD) in E. coli BL21(DE3) and successfully translocating it to the periplasmic region. This study represents the inaugural investigation into the utilisation of a naturally modified signal peptide, thereby corroborating the assertion that signal peptide deletion variants can influence protein secretion efficiency. Furthermore, the findings substantiate the proposition that such variants can serve as a viable alternative for the secretion of heterologous proteins in E. coli.

Reviews

Trans-acting regulators of ribonuclease activity: Jaejin Lee , Minho Lee , Kangseok Lee; J. Microbiol. 2021;59(4):341-359. Published online March 29, 2021; DOI: https://doi.org/10.1007/s12275-021-0650-6

57 View
0 Download
3 Web of Science
4 Crossref

Abstract

RNA metabolism needs to be tightly regulated in response to changes in cellular physiology. Ribonucleases (RNases) play an essential role in almost all aspects of RNA metabolism, including processing, degradation, and recycling of RNA molecules. Thus, living systems have evolved to regulate RNase activity at multiple levels, including transcription, post-transcription, post-translation, and cellular localization. In addition, various trans-acting regulators of RNase activity have been discovered in recent years. This review focuses on the physiological roles and underlying mechanisms of trans-acting regulators of RNase activity.

Citations

Citations to this article as recorded by

Comparative Transcriptomic Analysis of Flagellar-Associated Genes in Salmonella Typhimurium and Its rnc Mutant
Seungmok Han, Ji-Won Byun, Minho Lee
Journal of Microbiology.2024; 62(1): 33. CrossRef
Insights into the metabolism, signaling, and physiological effects of 2’,3’-cyclic nucleotide monophosphates in bacteria
Nick J. Marotta, Emily E. Weinert
Critical Reviews in Biochemistry and Molecular Biology.2023; 58(2-6): 118. CrossRef
Relaxed Cleavage Specificity of Hyperactive Variants of Escherichia coli RNase E on RNA I
Dayeong Bae, Hana Hyeon, Eunkyoung Shin, Ji-Hyun Yeom, Kangseok Lee
Journal of Microbiology.2023; 61(2): 211. CrossRef
Regulator of RNase E activity modulates the pathogenicity of Salmonella Typhimurium
Jaejin Lee, Eunkyoung Shin, Ji-Hyun Yeom, Jaeyoung Park, Sunwoo Kim, Minho Lee, Kangseok Lee
Microbial Pathogenesis.2022; 165: 105460. CrossRef

Ammonia-oxidizing archaea in biological interactions: Jong-Geol Kim , Khaled S. Gazi , Samuel Imisi Awala , Man-Young Jung , Sung-Keun Rhee; J. Microbiol. 2021;59(3):298-310. Published online February 23, 2021; DOI: https://doi.org/10.1007/s12275-021-1005-z

65 View
0 Download
14 Web of Science
15 Crossref

Abstract

The third domain Archaea was known to thrive in extreme or anoxic environments based on cultivation studies. Recent metagenomics- based approaches revealed a widespread abundance of archaea, including ammonia-oxidizing archaea (AOA) of Thaumarchaeota in non-extreme and oxic environments. AOA alter nitrogen species availability by mediating the first step of chemolithoautotrophic nitrification, ammonia oxidation to nitrite, and are important primary producers in ecosystems, which affects the distribution and activity of other organisms in ecosystems. Thus, information on the interactions of AOA with other cohabiting organisms is a crucial element in understanding nitrogen and carbon cycles in ecosystems as well as the functioning of whole ecosystems. AOA are self-nourishing, and thus interactions of AOA with other organisms can often be indirect and broad. Besides, there are possibilities of specific and obligate interactions. Mechanisms of interaction are often not clearly identified but only inferred due to limited knowledge on the interaction factors analyzed by current technologies. Here, we overviewed different types of AOA interactions with other cohabiting organisms, which contribute to understanding AOA functions in ecosystems.

Citations

Citations to this article as recorded by

Identification of structural and regulatory cell-shape determinants in Haloferax volcanii
Heather Schiller, Yirui Hong, Joshua Kouassi, Theopi Rados, Jasmin Kwak, Anthony DiLucido, Daniel Safer, Anita Marchfelder, Friedhelm Pfeiffer, Alexandre Bisson, Stefan Schulze, Mechthild Pohlschroder
Nature Communications.2024;[Epub] CrossRef
Nitrogen cycling process and application in different prawn culture modes
Zhao Chen, Jian Li, Qianqian Zhai, Zhiqiang Chang, Jitao Li
Reviews in Aquaculture.2024; 16(4): 1580. CrossRef
Multidrug-resistant plasmid RP4 inhibits the nitrogen removal capacity of ammonia-oxidizing archaea, ammonia-oxidizing bacteria, and comammox in activated sludge
Zhaohui Zhang, Lin Bo, Shang Wang, Chenyu Li, Xi Zhang, Bin Xue, Xiaobo Yang, Xinxin He, Zhiqiang Shen, Zhigang Qiu, Chen Zhao, Jingfeng Wang
Environmental Research.2024; 242: 117739. CrossRef
Distinct mechanisms drive plant-nitrifier interactions in topsoil and subsoil
Di Liang, Niuniu Ji, Angela Kent, Wendy H. Yang
Soil Biology and Biochemistry.2024; 192: 109370. CrossRef
Diversity, composition, metabolic characteristics, and assembly process of the microbial community in sewer system at the early stage
Yiming Yuan, Guangyi Zhang, Hongyuan Fang, Haifeng Guo, Yongkang Li, Zezhuang Li, Siwei Peng, Fuming Wang
Environmental Science and Pollution Research.2024; 31(9): 13075. CrossRef
Response of soil microbial community structure and function to the sewage leakage: A case study of a 25-year-old cesspool
Xiaocheng Wei, Jiayin Liang, Tianyang Ning, Chunxue Zhang, Jiarui Wang, Lu Tan, Feng Shen
Chemosphere.2024; 363: 142753. CrossRef
Hiding in plain sight: The discovery of complete genomes of 11 hypothetical spindle‐shaped viruses that putatively infect mesophilic ammonia‐oxidizing archaea
Yimin Ni, Tianqi Xu, Shuling Yan, Lanming Chen, Yongjie Wang
Environmental Microbiology Reports.2024;[Epub] CrossRef
Inulin from halophilic archaeon Haloarcula: Production, chemical characterization, biological, and technological properties
Alejandra Aragón-León, Lorena Moreno-Vilet, Marisela González-Ávila, Pedro Martín Mondragón-Cortez, Guilherme Lanzi Sassaki, Raúl Balam Martínez-Pérez, Rosa María Camacho-Ruíz
Carbohydrate Polymers.2023; 321: 121333. CrossRef
Uncovering the Prokaryotic Diversity of the Bathyal Waters above the Kuril–Kamchatka Trench
Susanna Gorrasi, Angelika Brandt, Francesca Pittino, Andrea Franzetti, Marcella Pasqualetti, Barbara Muñoz-Palazon, Giorgia Novello, Massimiliano Fenice
Journal of Marine Science and Engineering.2023; 11(11): 2145. CrossRef
Nitrous Oxide Distributions in the Oxygenated Water Column of the Sargasso Sea
Annaliese C. S. Meyer, Jay T. Cullen, Damian S. Grundle
Atmosphere-Ocean.2023; 61(3): 173. CrossRef
An Initial Proteomic Analysis of Biogas-Related Metabolism of Euryarchaeota Consortia in Sediments from the Santiago River, México
Jesús Barrera-Rojas, Kelly Joel Gurubel-Tun, Emmanuel Ríos-Castro, María Cristina López-Méndez, Belkis Sulbarán-Rangel
Microorganisms.2023; 11(7): 1640. CrossRef
Bacteria and Archaea Regulate Particulate Organic Matter Export in Suspended and Sinking Marine Particle Fractions
Choaro D. Dithugoe, Oliver K. I. Bezuidt, Emma L. Cavan, William P. Froneman, Sandy J. Thomalla, Thulani P. Makhalanyane, Barbara J. Campbell
mSphere.2023;[Epub] CrossRef
Insights into the prokaryotic communities of the abyssal-hadal benthic-boundary layer of the Kuril Kamchatka Trench
Susanna Gorrasi, Andrea Franzetti, Angelika Brandt, Ulrike Minzlaff, Marcella Pasqualetti, Massimiliano Fenice
Environmental Microbiome.2023;[Epub] CrossRef
Examining the Interaction Between Free‐Living Bacteria and Iron in the Global Ocean
Anh Le‐Duy Pham, Olivier Aumont, Lavenia Ratnarajah, Alessandro Tagliabue
Global Biogeochemical Cycles.2022;[Epub] CrossRef
Omics-based microbiome analysis in microbial ecology: from sequences to information
Jang-Cheon Cho
Journal of Microbiology.2021; 59(3): 229. CrossRef

Journal Article

Recombinant baculovirus-based vaccine expressing M2 protein induces protective CD8+ T-cell immunity against respiratory syncytial virus infection: Jeong-Yoon Lee , Jun Chang; J. Microbiol. 2017;55(11):900-908. Published online October 27, 2017; DOI: https://doi.org/10.1007/s12275-017-7306-6

57 View
0 Download
9 Crossref

Abstract

Respiratory syncytial virus (RSV) is an important cause of acute lower respiratory tract disease in infants, young children, immunocompromised individuals, and the elderly. However, despite ongoing efforts to develop an RSV vaccine, there is still no authorized RSV vaccine for humans. Baculovirus has attracted attention as a vaccine vector because of its ability to induce a high level of humoral and cellular immunity, low cytotoxicity against various antigens, and biological safety for humans. In this study, we constructed a recombinant baculovirus- based vaccine expressing the M2 protein of RSV under the control of cytomegalovirus promoter (Bac_RSVM2) to induce CD8+ T-cell responses which play an important role in viral clearance, and investigated its protective efficacy against RSV infection. Immunization with Bac_RSVM2 via intranasal or intramuscular route effectively elicited the specific CD8+ T-cell responses. Most notably, immunization with Bac_RSVM2 vaccine almost completely protected mice from RSV challenge without vaccine-enhanced immunopathology. In conclusion, these results suggest that Bac_RSVM2 vaccine employing the baculovirus delivery platform has promising potential to be developed as a safe and novel RSV vaccine that provides protection against RSV infection.

Citations

Citations to this article as recorded by

Respiratory delivered vaccines: Current status and perspectives in rational formulation design
Lan Wu, Wenwen Xu, Huiyang Jiang, Mingshi Yang, Dongmei Cun
Acta Pharmaceutica Sinica B.2024; 14(12): 5132. CrossRef
Enhanced virulence of genetically engineered Autographa californica nucleopolyhedrovirus owing to accelerated viral DNA replication aided by inserted ascovirus genes
Huan Yu, Chang-Jin Yang, Yi-Yi Ou-Yang, Yue Tong, Hui-Yu Lan, Jia-Min Gan, Shi-Wei Li, Ding-Yi Bai, Guo-Hua Huang
Pesticide Biochemistry and Physiology.2023; 192: 105382. CrossRef
Cytokines and CD8 T cell immunity during respiratory syncytial virus infection
Megan E. Schmidt, Steven M. Varga
Cytokine.2020; 133: 154481. CrossRef
Induction of mucosal immunity against pathogens by using recombinant baculoviral vectors: Mechanisms, advantages, and limitations
Mario Fragoso-Saavedra, Marco A Vega-López
Journal of Leukocyte Biology.2020; 108(3): 835. CrossRef
Endogenous n-3 Polyunsaturated Fatty Acids Are Beneficial to Dampen CD8+ T Cell-Mediated Inflammatory Response upon the Viral Infection in Mice
Kyung Won Kang, Seyoung Kim, Yong-Bin Cho, Seung Rok Ryu, Young-Jin Seo, Sang-Myeong Lee
International Journal of Molecular Sciences.2019; 20(18): 4510. CrossRef
Anti-viral activity of compounds from Agrimonia pilosa and Galla rhois extract mixture
Jeong Eun Kwon, Yeong-Geun Lee, Ji-Hun Kang, Yun-Feng Bai, Yong Joon Jeong, Nam-In Baek, Young-Jin Seo, Se Chan Kang
Bioorganic Chemistry.2019; 93: 103320. CrossRef
Vaccine containing G protein fragment and recombinant baculovirus expressing M2 protein induces protective immunity to respiratory syncytial virus
Yeong-Min Jo, Jungwoo Kim, Jun Chang
Clinical and Experimental Vaccine Research.2019; 8(1): 43. CrossRef
Recombinant live attenuated influenza vaccine viruses carrying CD8 T-cell epitopes of respiratory syncytial virus protect mice against both pathogens without inflammatory disease
Tatiana Kotomina, Irina Isakova-Sivak, Victoria Matyushenko, Ki-Hye Kim, Youri Lee, Yu-Jin Jung, Sang-Moo Kang, Larisa Rudenko
Antiviral Research.2019; 168: 9. CrossRef
The CD8 T Cell Response to Respiratory Virus Infections
Megan E. Schmidt, Steven M. Varga
Frontiers in Immunology.2018;[Epub] CrossRef

Review

MINIREVIEW] Advances in novel influenza vaccines: a patent review: Jae-Min Song; J. Microbiol. 2016;54(6):403-412. Published online May 27, 2016; DOI: https://doi.org/10.1007/s12275-016-6176-7

45 View
0 Download
1 Crossref

Abstract

The threat of a major human influenza pandemic such as the avian H5N1 or the 2009 new H1N1 has emphasized the need for effective prevention strategies to combat these pathogens. Although egg based influenza vaccines have been well established for a long time, it remains an ongoing public health need to develop alternative production methods that ensures improved safety, efficacy, and ease of administration compared with conventional influenza vaccines. This article is intended to cover some of the recent advances and related patents on the development of influenza vaccines including live attenuated, cell based, genomic and synthetic peptide vaccines.

Citations

Citations to this article as recorded by

Plant-made virus-like particles bearing influenza hemagglutinin (HA) recapitulate early interactions of native influenza virions with human monocytes/macrophages
Alexander I. Makarkov, Sabrina Chierzi, Stéphane Pillet, Keith K. Murai, Nathalie Landry, Brian J. Ward
Vaccine.2017; 35(35): 4629. CrossRef

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

Characterization of Recombinant β-Glucosidase from Arthrobacter chlorophenolicus and Biotransformation of Ginsenosides Rb1, Rb2, Rc, and Rd: Myung Keun Park , Chang-Hao Cui , Sung Chul Park , Seul-Ki Park , Jin-Kwang Kim , Mi-Sun Jung , Suk-Chae Jung , Mi-Sun Jung , Suk-Chae Jung , Sun-Chang Kim , Wan-Taek Im; J. Microbiol. 2014;52(5):399-406. Published online May 9, 2014; DOI: https://doi.org/10.1007/s12275-014-3601-7

53 View
0 Download
10 Crossref

Abstract

The focus of this study was the cloning, expression, and characterization of recombinant ginsenoside hydrolyzing β-glucosidase from Arthrobacter chlorophenolicus with an ultimate objective to more efficiently bio-transform ginse-nosides. The gene bglAch, consisting of 1,260 bp (419 amino acid residues) was cloned and the recombinant enzyme, over-expressed in Escherichia coli BL21 (DE3), was characterized. The GST-fused BglAch was purified using GST·Bind agarose resin and characterized. Under optimal conditions (pH 6.0 and 37°C) BglAch hydrolyzed the outer glucose and arabino-pyranose moieties of ginsenosides Rb1 and Rb2 at the C20 position of the aglycone into ginsenoside Rd. This was fol-lowed by hydrolysis into F2 of the outer glucose moiety of ginsenoside Rd at the C3 position of the aglycone. Additio-nally, BglAch more slowly transformed Rc to F2 via C-Mc1 (compared to hydrolysis of Rb1 or Rb2). These results in-dicate that the recombinant BglAch could be useful for the production of ginsenoside F2 for use in the pharmaceutical and cosmetic industries.

Citations

Citations to this article as recorded by

Production and pharmaceutical research of minor saponins in Panax notoginseng (Sanqi): Current status and future prospects
Hui Zhang, Jianxiu Li, Mengxue Diao, Jianbin Li, Nengzhong Xie
Phytochemistry.2024; 223: 114099. CrossRef
Microbial production and applications of β-glucosidase-A review
Wenqi Yang, Yaowu Su, Rubing Wang, Huanyu Zhang, Hongyan Jing, Jie Meng, Guoqi Zhang, Luqi Huang, Lanping Guo, Juan Wang, Wenyuan Gao
International Journal of Biological Macromolecules.2024; 256: 127915. CrossRef
Progress in the Conversion of Ginsenoside Rb1 into Minor Ginsenosides Using β-Glucosidases
Hongrong Zhu, Rui Zhang, Zunxi Huang, Junpei Zhou
Foods.2023; 12(2): 397. CrossRef
Enzymatic biotransformation of ginsenoside Rb1 by recombinant β-glucosidase of bacterial isolates from Indonesia
Almando Geraldi, Ni'matuzahroh, Fatimah, Chang-Hao Cui, Thi Thuy Nguyen, Sun Chang Kim
Biocatalysis and Agricultural Biotechnology.2020; 23: 101449. CrossRef
Characterization of a Novel Ginsenoside MT1 Produced by an Enzymatic Transrhamnosylation of Protopanaxatriol-Type Ginsenosides Re
Byeong-Min Jeon, Jong-In Baek, Min-Sung Kim, Sun-Chang Kim, Chang-hao Cui
Biomolecules.2020; 10(4): 525. CrossRef
In silico Approach to Elucidate Factors Associated with GH1 β-Glucosidase Thermostability
Amer Ahmed, Ayesha Sumreen, Aasia Bibi, Faiz ul Hassan Nasim, Kashfa Batool
Journal of Pure and Applied Microbiology.2019; 13(4): 1953. CrossRef
A literature update elucidating production of Panax ginsenosides with a special focus on strategies enriching the anti-neoplastic minor ginsenosides in ginseng preparations
Tanya Biswas, A. K. Mathur, Archana Mathur
Applied Microbiology and Biotechnology.2017; 101(10): 4009. CrossRef
Classification of glycosidases that hydrolyze the specific positions and types of sugar moieties in ginsenosides
Kyung-Chul Shin, Deok-Kun Oh
Critical Reviews in Biotechnology.2016; 36(6): 1036. CrossRef
Insight into a novel β-1,4-glucosidase from Streptomyces griseorubens JSD-1
H.-W. Feng, Y.-E. Zhi, Y.-J. Sun, L.-R. Xu, L.-M. Wang, X.-J. Zhan, P. Zhou
Applied Biochemistry and Microbiology.2016; 52(4): 371. CrossRef
Overexpression and characterization of a glycoside hydrolase family 1 enzyme from Cellulosimicrobium cellulans sp. 21 and its application for minor ginsenosides production
Ye Yuan, Yanbo Hu, Chenxing Hu, Jiayi Leng, Honglei Chen, Xuesong Zhao, Juan Gao, Yifa Zhou
Journal of Molecular Catalysis B: Enzymatic.2015; 120: 60. CrossRef

Low-Scale Expression and Purification of an Active Putative Iduronate 2-Sulfate Sulfatase-Like Enzyme from Escherichia coli K12: Edwin David Morales-Álvarez , Claudia Marcela Rivera-Hoyos , Angélica María Baena-Moncada , Patricia Landázuri , Raúl A. Poutou-Piñales , Homero Sáenz-Suárez , Luis A. Barrera , Olga Y. Echeverri-Peña; J. Microbiol. 2013;51(2):213-221. Published online April 27, 2013; DOI: https://doi.org/10.1007/s12275-013-2416-2

44 View
0 Download
15 Scopus

Abstract: The sulfatase family involves a group of enzymes with a large degree of similarity. Until now, sixteen human sulfatases have been identified, most of them found in lysosomes. Human deficiency of sulfatases generates various genetic disorders characterized by abnormal accumulation of sulfated intermediate compounds. Mucopolysaccharidosis type II is characterized by the deficiency of iduronate 2-sulfate sulfatase (IDS), causing the lysosomal accumulation of heparan and dermatan sulfates. Currently, there are several cases of genetic diseases treated with enzyme replacement therapy, which have generated a great interest in the development of systems for recombinant protein expression. In this work we expressed the human recombinant IDS-Like enzyme (hrIDS-Like) in Escherichia coli DH5α. The enzyme concentration revealed by ELISA varied from 78.13 to 94.35 ng/ml and the specific activity varied from 34.20 to 25.97 nmol/h/mg. Western blotting done after affinity chromatography purification showed a single band of approximately 40 kDa, which was recognized by an IgY polyclonal antibody that was developed against the specific peptide of the native protein. Our 100 ml-shake-flask assays allowed us to improve the enzyme activity seven fold, compared to the E. coli JM109/pUC13-hrIDS-Like system. Additionally, the results obtained in the present study were equal to those obtained with the Pichia pastoris GS1115/ pPIC-9-hrIDS-Like system (3 L bioreactor scale). The system used in this work (E. coli DH5α/pGEX-3X-hrIDS-Like) emerges as a strategy for improving protein expression and purification, aimed at recombinant protein chemical characterization, future laboratory assays for enzyme replacement therapy, and as new evidence of active putative sulfatase production in E. coli.

NOTE] Biosynthetic Pathway for Poly(3-Hydroxypropionate) in Recombinant Escherichia coli: Qi Wang , Changshui Liu , Mo Xian , Yongguang Zhang , Guang Zhao; J. Microbiol. 2012;50(4):693-697. Published online August 25, 2012; DOI: https://doi.org/10.1007/s12275-012-2234-y

38 View
0 Download
38 Scopus

Abstract: Poly(3-hydroxypropionate) (P3HP) is a biodegradable and biocompatible thermoplastic. In this study, we engineered a P3HP biosynthetic pathway in recombinant Escherichia coli. The genes for malonyl-CoA reductase (mcr, from Chloroflexus aurantiacus), propionyl-CoA synthetase (prpE, from E. coli), and polyhydroxyalkanoate synthase (phaC1, from Ralstonia eutropha) were cloned and expressed in E. coli. The E. coli genes accABCD encoding acetyl-CoA carboxylase were used to channel the carbon into the P3HP pathway. Using glucose as a sole carbon source, the cell yield and P3HP content were 1.32 g/L and 0.98% (wt/wt [cell dry weight]), respectively. Although the yield is relatively low, our study shows the feasibility of engineering a P3HP biosynthetic pathway using a structurally unrelated carbon source in bacteria.

Expression and Purification of Lacticin Q by Small Ubiquitin-Related Modifier Fusion in Escherichia coli: Qingshan Ma , Zhanqiao Yu , Bing Han , Qing Wang , Rijun Zhang; J. Microbiol. 2012;50(2):326-331. Published online April 27, 2012; DOI: https://doi.org/10.1007/s12275-012-1425-x

29 View
0 Download
13 Scopus

Abstract: Lacticin Q is a broad-spectrum class II bacteriocin with potential as an alternative to conventional antibiotics. The objective of this study was to produce recombinant lacticin Q using a small ubiquitin-related modifier (SUMO) fusion protein expression system. The 168-bp lacticin Q gene was cloned into the expression vector pET SUMO and transformed into Escherichia coli BL21(DE3). The soluble fusion protein was recovered with a Ni-NTA Sepharose column (95% purity); 130 mg protein was obtained per liter of fermentation culture. The SUMO tag was then proteolytically cleaved from the protein, which was re-applied to the column. Finally, about 32 mg lacticin Q (≥96% purity) was obtained. The recombinant protein exhibited antimicrobial properties similar to that of the native protein, demonstrating that lacticin Q had been successfully expressed by the SUMO fusion system.

NOTE] Development of a High-Throughput Screening Method for Recombinant Escherichia coli with Intracellular Dextransucrase Activity: So-Ra Lee , Ah-Rum Yi , Hong-Gyun Lee , Myoung-Uoon Jang , Jung-Mi Park , Nam Soo Han , Tae-Jip Kim; J. Microbiol. 2011;49(2):320-323. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-1078-1

33 View
0 Download
1 Scopus

Abstract: To efficiently engineer intracellular dextransucrase (DSase) expression in Escherichia coli, a high-throughput screening method was developed based on the polymer-forming activity of the enzyme. Recombinant E. coli containing the Leuconostoc citreum DSase (LcDS) gene was grown on Luria-Bertani agar plates, containing 2% sucrose, at 37°C for 8 h. The plates were then evenly overlaid with 0.6% soft agar, containing 1.2 mg/ml D-cycloserine, and incubated at 30°C to allow gradual cell disruption until a dextran polymer grew through the overlaid layer. A significant correlation between dextran size and enzyme activity was established and applied for screening truncated mutants with LcDS activity.

Retracted Publication

Identification and Characterization of a Novel Antibacterial Peptide, Avian β-Defensin 2 from Ducks: Deying Ma , Ruiqin Wang , Wenyan Liao , Zongxi Han , Shengwang Liu; J. Microbiol. 2009;47(5):610-618. Published online October 24, 2009; DOI: https://doi.org/10.1007/s12275-009-0068-z

38 View
0 Download
33 Scopus

Abstract: In this study, a novel avian β-defensin (AvBD) was isolated from duck pancreas. The complete nucleotide sequence of the gene contained an 195 bp open reading frame encoding 64 amino acids. Homology, characterization and comparison of the gene with AvBD from other avian species confirmed that it was duck AvBD2. The mRNA expression of the gene was analyzed in 17 tissues from 21-day-old ducks. AvBD2 was highly expressed in the trachea, crop, heart, bone marrow, and pancreas; moderately expressed in the muscular stomach, small intestine, kidney, spleen, thymus, and bursa of Fabricius; and weakly expressed in skin. We produced and purified recombinant AvBD2 by expressing the gene in Escherichia coli. As expected, the recombinant peptide exhibited strong bactericidal properties against Bacillus cereus, Staphylococcus aureus, and Pasteurella multocida, and weak bactericidal properties against E. coli and Salmonella choleraesuis. In addition, the recombinant protein retained antimicrobial activity against S. aureus under different temperatures (range, -20°C to 100°C) and pH values (range, 3 to 12).

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

Stable Expression and Secretion of Polyhydroxybutyrate Depolymerase of Paucimonas lemoignei in Escherichia coli: Se Whan Park , Moon Gyu Chung , Hwa Young Lee , Jeong Yoon Kim , Young Ha Rhee; J. Microbiol. 2008;46(6):662-669. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0283-z

43 View
0 Download
1 Scopus

Abstract: An efficient strategy for the expression and secretion of extracellular polyhydroxybutyrate depolymerase (PhaZ1) of Paucimonas lemoignei in Escherichia coli was developed by employing the signal peptide of PhaZ1 and a truncated ice nucleation protein anchoring motif (INPNC). Directly synthesized mature form of PhaZ1 was present in the cytoplasm of host cells as inclusion bodies, while a construct containing PhaZ1 and its own N-terminal signal peptide (PrePhaZ1) enabled the secretion of active PhaZ1 into the extracellular medium. However, the PrePhaZ1 construct was harmful to the host cell and resulted in atypical growth and instability of the plasmid during the cultivation. In contrast, INPNC-PhaZ1 and INPNCPrePhaZ1 fusion constructs did not affect growth of host cells. INPNC-PhaZ1 was successfully displayed on the cell surface with its fusion form, but did not retain PhaZ1 activity. In the case of INPNC-PrePhaZ1, the initially synthesized fusion form was separated by precise cleavage of the signal peptide, and active PhaZ1 was consequently released into the culture medium. The amount of PhaZ1 derived from E. coli (INPNC-PrePhaZ1) was almost twice as great as that directly expressed from E. coli (PrePhaZ1), and was predominantly (approximately 85%) located in the periplasm when cultivated at 22°C but was efficiently secreted into the extracellular medium when cultivated at 37°C.

Journal Articles

Biologically Active and C-Amidated HinnavinII-38-Asn Produced from a Trx Fusion Construct in Escherichia coli: Chang Soo Kang , Seung-Yeol Son , In Seok Bang; J. Microbiol. 2008;46(6):656-661. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0214-z

42 View
0 Download
9 Scopus

Abstract: The cabbage butterfly (Artogeia rapae) antimicrobial peptide hinnavinII as a member of cecropin family is synthesized as 37 residues in size with an amidated lysine at C-terminus and shows the humoral immune response to a bacterial invasion. In this work, a synthetic gene for hinnavinII-38-Asn (HIN) with an additional amino acid asparagine residue containing amide group at C-terminus was cloned into pET-32a(+) vector to allow expression of HIN as a Trx fusion protein in Escherichia coli strain BL21 (DE3) pLysS. The resulting expression level of the fusion protein Trx-HIN could reach 15~20% of the total cell proteins and more than 70% of the target proteins were in soluble form. The fusion protein could be purified successfully by HiTrap Chelating HP column and a high yield of 15 mg purified fusion protein was obtained from 80 ml E. coli culture. Recombinant HIN was readily obtained by enterokinase cleavage of the fusion protein followed by FPLC chromatography, and 3.18 mg pure active recombinant HIN was obtained from 80 ml culture. The molecular mass of recombinant HIN determined by MALDI-TOF mass spectrometer is 4252.084 Da which matches the theoretical mass (4252.0 Da) of HIN. Comparing the antimicrobial activities of the recombinant hinnavinII with C-amidated terminus to that without an amidated C-terminus, we found that the amide of asparagine at C-terminus of hinnavinII improved its potency on certain microorganism such as E. coli, Enterobacter cloacae, Bacillus megaterium, and Staphylococcus aureus.

Characterization of the Bacillus subtilis WL-3 Mannanase from a Recombinant Escherichia coli: Ki-Hong Yoon , Seesub Chung , Byung-Lak Lim; J. Microbiol. 2008;46(3):344-349. Published online July 5, 2008; DOI: https://doi.org/10.1007/s12275-008-0045-y

35 View
0 Download
31 Scopus

Abstract: A mannanase was purified from a cell-free extract of the recombinant Escherichia coli carrying a Bacillus subtilis WL-3 mannanase gene. The molecular mass of the purified mannanase was 38 kDa as estimated by SDS-PAGE. Optimal conditions for the purified enzyme occurred at pH 6.0 and 60°C. The specific activity of the purified mannanase was 5,900 U/mg on locust bean gum (LBG) galactomannan at pH 6.0 and 50°C. The activity of the enzyme was slightly inhibited by Mg2+, Ca2+, EDTA and SDS, and noticeably enhanced by Fe2+. When the enzyme was incubated at 4°C for one day in the presence of 3 mM Fe2+, no residual activity of the mannanase was observed. The enzyme showed higher activity on LBG and konjac glucomannan than on guar gum galactomannan. Furthermore, it could hydrolyze xylans such as arabinoxylan, birchwood xylan and oat spelt xylan, while it did not exhibit any activities towards carboxymethylcellulose and para-nitrophenyl-β-mannopyranoside. The predominant products resulting from the mannanase hydrolysis were mannose, mannobiose and mannotriose for LBG or mannooligosaccharides including mannotriose, mannotetraose, mannopentaose and mannohexaose. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose.

First
Prev
Page of 2
Next
Last

Search