Journal of Microbiology

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CA‑CAS‑01‑A: A Permissive Cell Line for Isolation and Live Attenuated Vaccine Development Against African Swine Fever Virus: Seung-Chul Lee , Yongkwan Kim , Ji-Won Cha , Kiramage Chathuranga , Niranjan Dodantenna , Hyeok-Il Kwon , Min Ho Kim , Weonhwa Jheong , In-Joong Yoon , Joo Young Lee , Sung-Sik Yoo , Jong-Soo Lee; J. Microbiol. 2024;62(2):125-134. Published online March 13, 2024; DOI: https://doi.org/10.1007/s12275-024-00116-1

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African swine fever virus (ASFV) is the causative agent of the highly lethal African swine fever disease that affects domestic pigs and wild boars. In spite of the rapid spread of the virus worldwide, there is no licensed vaccine available. The lack of a suitable cell line for ASFV propagation hinders the development of a safe and effective vaccine. For ASFV propagation, primary swine macrophages and monocytes have been widely studied. However, obtaining these cells can be time-consuming and expensive, making them unsuitable for mass vaccine production. The goal of this study was to validate the suitability of novel CA-CAS-01-A (CAS-01) cells, which was identified as a highly permissive cell clone for ASFV replication in the MA-104 parental cell line for live attenuated vaccine development. Through a screening experiment, maximum ASFV replication was observed in the CAS-01 cell compared to other sub-clones of MA-104 with 14.89 and log10 7.5 ± 0.15 Ct value and TCID50/ ml value respectively. When CAS-01 cells are inoculated with ASFV, replication of ASFV was confirmed by Ct value for ASFV DNA, HAD50/ ml assay, TCID50/ ml assay, and cytopathic effects and hemadsoption were observed similar to those in primary porcine alveolar macrophages after 5th passage. Additionally, we demonstrated stable replication and adaptation of ASFV over the serial passage. These results suggest that CAS-01 cells will be a valuable and promising cell line for ASFV isolation, replication, and development of live attenuated vaccines.

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Development and characterization of high-efficiency cell-adapted live attenuated vaccine candidate against African swine fever
Min Ho Kim, Ashan Subasinghe, Yongkwan Kim, Hyeok-Il Kwon, Yehjin Cho, Kiramage Chathuranga, Ji-Won Cha, Ji-Yoon Moon, Ji-Hyeon Hong, Jin Kim, Seung-Chul Lee, Niranjan Dodantenna, Nuwan Gamage, W. A. Gayan Chathuranga, Yeonji Kim, In-Joong Yoon, Joo Young
Emerging Microbes & Infections.2024;[Epub] CrossRef

Analysis of the L-malate biosynthesis pathway involved in poly(β-L-malic acid) production in Aureobasidium melanogenum GXZ-6 by addition of metabolic intermediates and inhibitors: Wei Zeng , Bin Zhang , Qi Liu , Guiguang Chen , Zhiqun Liang; J. Microbiol. 2019;57(4):281-287. Published online February 5, 2019; DOI: https://doi.org/10.1007/s12275-019-8424-0

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Abstract

Poly(β-L-malic acid) (PMA) is a promising polyester formed from L-malate in microbial cells. Malate biosynthesis is crucial for PMA production. Previous studies have shown that the non-oxidative pathway or oxidative pathway (TCA cycle) is the main biosynthetic pathway of malate in most of PMAproducing strains, while the glyoxylate cycle is only a supplementary pathway. In this study, we investigated the effect of exogenous metabolic intermediates and inhibitors of the malate biosynthetic pathway on PMA production by Aureobasidium melanogenum GXZ-6. The results showed that PMA production was stimulated by maleic acid (a fumarase inhibitor) and sodium malonate (a succinate dehydrogenase inhibitor) but inhibited by succinic acid and fumaric acid. This indicated that the TCA cycle might not be the only biosynthetic pathway of malate. In addition, the PMA titer increased by 18.1% upon the addition of glyoxylic acid after 72 h of fermentation, but the PMA titer decreased by 7.5% when itaconic acid (an isocitrate lyase inhibitor) was added, which indicated that malate for PMA production was synthesized significantly via the glyoxylate cycle rather than the TCA cycle. Furthermore, in vitro enzyme activities of the TCA and glyoxylate cycles suggested that the glyoxylate cycle significantly contributed to the PMA production, which is contradictory to what has been reported previously in other PMA-producing A. pullulans.

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De novo transcriptome assembly of Aureobasidium melanogenum CGMCC18996 to analyze the β-poly(L-malic acid) biosynthesis pathway under the CaCO3 addition
Genan Wang, Haisong Yin, Tingbin Zhao, Donglin Yang, Shiru Jia, Changsheng Qiao
Food Science and Human Wellness.2023; 12(4): 1248. CrossRef
Microbial L-malic acid production: History, current progress, and perspectives
Yongyan Xi, Feiyu Fan, Xueli Zhang
Green Carbon.2023; 1(2): 118. CrossRef
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Bioprocess and Biosystems Engineering.2022; 45(10): 1673. CrossRef
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Frontiers in Microbiology.2021;[Epub] CrossRef
Small-Sized Co-Polymers for Targeted Delivery of Multiple Imaging and Therapeutic Agents
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Trends in Biochemical Sciences.2021; 46(3): 213. CrossRef
Polymalate (PMA) biosynthesis and its molecular regulation in Aureobasidium spp.
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International Journal of Biological Macromolecules.2021; 174: 512. CrossRef
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Thi Bich Huong Duong, Prattana Ketbot, Paripok Phitsuwan, Rattiya Waeonukul, Chakrit Tachaapaikoon, Akihiko Kosugi, Khanok Ratanakhanokchai, Patthra Pason
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FEMS Yeast Research.2020;[Epub] CrossRef
Effects of corn steep liquor on β-poly(l-malic acid) production in Aureobasidium melanogenum
Genan Wang, Bingyi Shi, Pan Zhang, Tingbin Zhao, Haisong Yin, Changsheng Qiao
AMB Express.2020;[Epub] CrossRef
A novel PMA synthetase is the key enzyme for polymalate biosynthesis and its gene is regulated by a calcium signaling pathway in Aureobasidium melanogenum ATCC62921
Kai Wang, Zhe Chi, Guang-Lei Liu, Cong-Yan Qi, Hong Jiang, Zhong Hu, Zhen-Ming Chi
International Journal of Biological Macromolecules.2020; 156: 1053. CrossRef
Poly(malic acid) production from liquefied corn starch by simultaneous saccharification and fermentation with a novel isolated Aureobasidium pullulans GXL-1 strain and its techno-economic analysis
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Bioresource Technology.2020; 304: 122990. CrossRef
Recent progress on bio-based production of dicarboxylic acids in yeast
Xi Zhang, Yunying Zhao, Yingli Liu, Jing Wang, Yu Deng
Applied Microbiology and Biotechnology.2020; 104(10): 4259. CrossRef

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