Journal of Microbiology

Journal Articles

Development of a Novel D‑Lactic Acid Production Platform Based on Lactobacillus saerimneri TBRC 5746: Kitisak Sansatchanon , Pipat Sudying , Peerada Promdonkoy , Yutthana Kingcha , Wonnop Visessanguan , Sutipa Tanapongpipat , Weerawat Runguphan , Kanokarn Kocharin; J. Microbiol. 2023;61(9):853-863. Published online September 14, 2023; DOI: https://doi.org/10.1007/s12275-023-00077-x

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D-Lactic acid is a chiral, three-carbon organic acid, that bolsters the thermostability of polylactic acid. In this study, we developed a microbial production platform for the high-titer production of D-lactic acid. We screened 600 isolates of lactic acid bacteria (LAB) and identified twelve strains that exclusively produced D-lactic acid in high titers. Of these strains, Lactobacillus saerimneri TBRC 5746 was selected for further development because of its homofermentative metabolism. We investigated the effects of high temperature and the use of cheap, renewable carbon sources on lactic acid production and observed a titer of 99.4 g/L and a yield of 0.90 g/g glucose (90% of the theoretical yield). However, we also observed L-lactic acid production, which reduced the product’s optical purity. We then used CRISPR/dCas9-assisted transcriptional repression to repress the two Lldh genes in the genome of L. saerimneri TBRC 5746, resulting in a 38% increase in D-lactic acid production and an improvement in optical purity. This is the first demonstration of CRISPR/dCas9-assisted transcriptional repression in this microbial host and represents progress toward efficient microbial production of D-lactic acid.

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Industrial–scale production of various bio–commodities by engineered microbial cell factories: Strategies of engineering in microbial robustness
Ju-Hyeong Jung, Vinoth Kumar Ponnusamy, Gopalakrishnan Kumar, Bartłomiej Igliński, Vinod Kumar, Grzegorz Piechota
Chemical Engineering Journal.2024; 502: 157679. CrossRef
Microbial Cell Factories: Biodiversity, Pathway Construction, Robustness, and Industrial Applicability
Rida Chaudhary, Ali Nawaz, Mireille Fouillaud, Laurent Dufossé, Ikram ul Haq, Hamid Mukhtar
Microbiology Research.2024; 15(1): 247. CrossRef
Adaptive Evolution for the Efficient Production of High-Quality d-Lactic Acid Using Engineered Klebsiella pneumoniae
Bo Jiang, Jiezheng Liu, Jingnan Wang, Guang Zhao, Zhe Zhao
Microorganisms.2024; 12(6): 1167. CrossRef
Enhancing D-lactic acid production from non-detoxified corn stover hydrolysate via innovative F127-IEA hydrogel-mediated immobilization of Lactobacillus bulgaricus T15
Yuhan Zheng, Feiyang Sun, Siyi Liu, Gang Wang, Huan Chen, Yongxin Guo, Xiufeng Wang, Maia Lia Escobar Bonora, Sitong Zhang, Yanli Li, Guang Chen
Frontiers in Microbiology.2024;[Epub] CrossRef

Analysis of a bac operon-silenced strain suggests pleiotropic effects of bacilysin in Bacillus subtilis: Ozan Ertekin , Meltem Kutnu , Aslı Aras Ta&# , Mustafa Demir , Ayten Yazgan Karata&# , Gülay Özcengiz; J. Microbiol. 2020;58(4):297-313. Published online January 28, 2020; DOI: https://doi.org/10.1007/s12275-020-9064-0

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Abstract

Bacilysin, as the simplest peptide antibiotic made up of only L-alanine and L-anticapsin, is produced and excreted by Bacillus subtilis under the control of quorum sensing. We analyzed bacilysin-nonproducing strain OGU1 which was obtained by bacA-targeted pMutin T3 insertion into the parental strain genome resulting in a genomic organization (bacA􍿁::lacZ::erm::bacABCDEF) to form an IPTG-inducible bac operon. Although IPTG induction provided 3- to 5-fold increment in the transcription of bac operon genes, no bacilysin activity was detectable in bioassays and inability of the OGU1 to form bacilysin was confirmed by UPLC-mass spectrometry analysis. Phenotypic analyses revealed the deficiencies in OGU1 with respect to colony pigmentation, spore coat proteins, spore resistance and germination, which could be rescued by external addition of bacilysin concentrate into its cultures. 2DE MALDI-TOF/MS and nanoLC-MS/MS were used as complementary approaches to compare cytosolic proteomes of OGU1. 2-DE identified 159 differentially expressed proteins corresponding to 121 distinct ORFs. In nanoLCMS/ MS, 76 proteins were differentially expressed in OGU1. Quantitative transcript analyses of selected genes validated the proteomic findings. Overall, the results pointed to the impact of bacilysin on expression of certain proteins of sporulation and morphogenesis; the members of mother cell compartment- specific σE and σK regulons in particular, quorum sensing and two component-global regulatory systems, peptide transport, stress response as well as CodY- and ScoCregulated proteins.

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Biocontrol Ability of Strain Bacillus amyloliquefaciens SQ-2 against Table Grape Rot Caused by Aspergillus tubingensis
Suran Li, Shuangshuang Dai, Lei Huang, Yumeng Cui, Ming Ying
Journal of Agricultural and Food Chemistry.2024; 72(44): 24374. CrossRef
Isolation and identification of a novel Bacillus velezensis strain JIN4 and its potential for biocontrol of kiwifruit bacterial canker caused by Pseudomonas syringae pv. actinidiae
Xin Zhao, Yang Zhai, Lin Wei, Fei Xia, Yuanru Yang, Yongjian Yi, Hongying Wang, Caisheng Qiu, Feng Wang, Liangbin Zeng
Frontiers in Plant Science.2024;[Epub] CrossRef
Signatures of kin selection in a natural population of the bacteria Bacillus subtilis
Laurence J Belcher, Anna E Dewar, Chunhui Hao, Melanie Ghoul, Stuart A West
Evolution Letters.2023; 7(5): 315. CrossRef
Comparative biological network analysis for differentially expressed proteins as a function of bacilysin biosynthesis in Bacillus subtilis
Meltem Kutnu, Elif Tekin İşlerel, Nurcan Tunçbağ, Gülay Özcengiz
Integrative Biology.2022; 14(5): 99. CrossRef
Probiotic effects of the Bacillus velezensis GY65 strain in the mandarin fish, Siniperca chuatsi
Jiachuan Wang, Defeng Zhang, Yajun Wang, Zhijun Liu, Lijuan Liu, Cunbin Shi
Aquaculture Reports.2021; 21: 100902. CrossRef
Bacilysin within the Bacillus subtilis group: gene prevalence versus antagonistic activity against Gram-negative foodborne pathogens
Catherine Nannan, Huong Quynh Vu, Annika Gillis, Simon Caulier, Thuy Thanh Thi Nguyen, Jacques Mahillon
Journal of Biotechnology.2021; 327: 28. CrossRef
Impact of spatial proximity on territoriality among human skin bacteria
Jhonatan A. Hernandez-Valdes, Lu Zhou, Marcel P. de Vries, Oscar P. Kuipers
npj Biofilms and Microbiomes.2020;[Epub] CrossRef

Microbial diversity in the rumen, reticulum, omasum, and abomasum of yak on a rapid fattening regime in an agro-pastoral transition zone: Dan Xue , Huai Chen , Xiaolin Luo , Jiuqiang Guan , Yixin He , Xinquan Zhao; J. Microbiol. 2018;56(10):734-743. Published online August 22, 2018; DOI: https://doi.org/10.1007/s12275-018-8133-0

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Abstract

The ruminant digestive system harbors a complex gut microbiome, which is poorly understood in the case of the four stomach compartments of yak. High-throughput sequencing and quantitative PCR were used to analyse microbial communities in the rumen, reticulum, omasum, and abomasum of six domesticated yak. The diversity of prokaryotes was higher in reticulum and omasum than in rumen and abomasum. Bacteroidetes predominated in the four stomach compartments, with abundance gradually decreasing in the trend rumen > reticulum > omasum > abomasum. Microorganism composition was different among the four compartments, all of which contained high levels of bacteria, methanogens, protozoa and anaerobic fungi. Some prokaryotic genera were associated with volatile fatty acids and pH. This study provides the first insights into the microorganism composition of four stomach compartments in yak, and may provide a foundation for future studies in this area.

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Pengujian Potensi Cairan Omasum Sapi untuk Ketercernaan Tiga Jenis Bahan Organik sebagai Sumber Bioetanol Generasi Ke-2
Endah Dwi Hastuti, Riska Amalia, Munifatul Izzati
Buletin Anatomi dan Fisiologi.2024; 9(1): 75. CrossRef
Effects of Two Feeding Patterns on Growth Performance, Rumen Fermentation Parameters, and Bacterial Community Composition in Yak Calves
Qin Li, Yan Tu, Tao Ma, Kai Cui, Jianxin Zhang, Qiyu Diao, Yanliang Bi
Microorganisms.2023; 11(3): 576. CrossRef
Transcriptome profiling in rumen, reticulum, omasum, and abomasum tissues during the developmental transition of pre-ruminant to the ruminant in yaks
Yili Liu, Qi Min, Jiao Tang, Lu Yang, Xinxin Meng, Tao Peng, Mingfeng Jiang
Frontiers in Veterinary Science.2023;[Epub] CrossRef
Nutrient availability of roughages in isocaloric and isonitrogenous diets alters the bacterial networks in the whole gastrointestinal tract of Hu sheep
Yuqi Li, Jian Gao, Yihan Xue, Ruolin Sun, Xiaoni Sun, Zhanying Sun, Suozhu Liu, Zhankun Tan, Weiyun Zhu, Yanfen Cheng
BMC Microbiology.2023;[Epub] CrossRef
Review: Effect of Experimental Diets on the Microbiome of Productive Animals
Rodrigo Huaiquipán, John Quiñones, Rommy Díaz, Carla Velásquez, Gastón Sepúlveda, Lidiana Velázquez, Erwin A. Paz, Daniela Tapia, David Cancino, Néstor Sepúlveda
Microorganisms.2023; 11(9): 2219. CrossRef
“The Yak”—A remarkable animal living in a harsh environment: An overview of its feeding, growth, production performance, and contribution to food security
Ali Mujtaba Shah, Iqra Bano, Izhar Hyder Qazi, Maharach Matra, Metha Wanapat
Frontiers in Veterinary Science.2023;[Epub] CrossRef
Effects of Two Different Straw Pellets on Yak Growth Performance and Ruminal Microbiota during Cold Season
Xiangyan Wang, Bingang Shi, Zhi Zuo, Youpeng Qi, Shijie Zhao, Xueping Zhang, Lijuan Lan, Yu Shi, Xiu Liu, Shaobin Li, Jiqing Wang, Jiang Hu
Animals.2023; 13(3): 335. CrossRef
Isoacids supplementation improves growth performance and feed fiber digestibility associated with ruminal bacterial community in yaks
Fei Jiang, Yanhua Gao, Zhongli Peng, Xiulian Ma, Yinjie You, Zhibin Hu, Anxiang He, Yupeng Liao
Frontiers in Microbiology.2023;[Epub] CrossRef
Representativeness of Fecal Microbiota Is Limited to Cecum and Colon in Domestic Yak
Wen Qin, Pengfei Song, Jirong Li, Jiuxiang Xie, Shoudong Zhang
Sustainability.2022; 14(16): 10263. CrossRef
Dynamic changes in the yak rumen eukaryotic community and metabolome characteristics in response to feed type
Xiaojing Cui, Yue Liu, Hao Wu, Qingxiang Meng, Shujie Liu, Shatuo Chai, Lizhuang Hao, Zhenming Zhou
Frontiers in Veterinary Science.2022;[Epub] CrossRef
The Response of Ruminal Microbiota and Metabolites to Different Dietary Protein Levels in Tibetan Sheep on the Qinghai-Tibetan Plateau
Xungang Wang, Tianwei Xu, Xiaoling Zhang, Na Zhao, Linyong Hu, Hongjin Liu, Qian Zhang, Yuanyue Geng, Shengping Kang, Shixiao Xu
Frontiers in Veterinary Science.2022;[Epub] CrossRef
Yak Gut Microbiota: A Systematic Review and Meta-Analysis
Yuxin Su, Junhong Su, Fanglin Li, Xiaojing Tian, Zewen Liu, Gongtao Ding, Jialin Bai, Zhuo Li, Zhongren Ma, Maikel P. Peppelenbosch
Frontiers in Veterinary Science.2022;[Epub] CrossRef
Gastrointestinal Biogeography of Luminal Microbiota and Short-Chain Fatty Acids in Sika Deer (Cervus nippon)
Xiaolong Hu, Yuting Wei, Tianxiang Zhang, Xiaoguo Wang, Yongtao Xu, Weiwei Zhang, Yunlin Zheng, Martha Vives
Applied and Environmental Microbiology.2022;[Epub] CrossRef
Comparison of Microbial Community and Metabolites in Four Stomach Compartments of Myostatin-Gene-Edited and Non-edited Cattle
Xinyu Zhou, Mingjuan Gu, Lin Zhu, Di Wu, Miaomiao Yang, Yajie Gao, Xueqiao Wang, Chunling Bai, Zhuying Wei, Lei Yang, Guangpeng Li
Frontiers in Microbiology.2022;[Epub] CrossRef
Effects of trace minerals supply from rumen sustained release boluses on milk yields and components, rumen fermentation and the rumen bacteria in lactating yaks (Bos grunniens)
Z.W. Zhao, Z.Y. Ma, H.C. Wang, C.F. Zhang
Animal Feed Science and Technology.2022; 283: 115184. CrossRef
Characterization of the bacterial microbiota across the different intestinal segments of the Qinghai semi-fine wool sheep on the Qinghai-Tibetan Plateau
Xungang Wang, Linyong Hu, Hongjin Liu, Tianwei Xu, Na Zhao, Xiaoling Zhang, Yuanyue Geng, Shengping Kang, Shixiao Xu
Animal Bioscience.2021; 34(12): 1921. CrossRef
Multi-Omics Analysis Reveals a Dependent Relationship Between Rumen Bacteria and Diet of Grass- and Grain-Fed Yaks
Chenchen Xu, Wenwen Liu, Baozhong Sun, Songshan Zhang, Shou Zhang, Yuanli Yang, Yuanhua Lei, Lan Chang, Peng Xie, Huayi Suo
Frontiers in Microbiology.2021;[Epub] CrossRef
Bacterial Community Characteristics in the Gastrointestinal Tract of Yak (Bos grunniens) Fully Grazed on Pasture of the Qinghai-Tibetan Plateau of China
Xueping Han, Hongjin Liu, Linyong Hu, Na Zhao, Shixiao Xu, Zhijia Lin, Yongwei Chen
Animals.2021; 11(8): 2243. CrossRef
Effects of rumen-protected methionine and lysine supplementation on milk yields and components, rumen fermentation, and the rumen microbiome in lactating yaks (Bos grunniens)
Z.W. Zhao, Z.Y. Ma, H.C. Wang, C.F. Zhang
Animal Feed Science and Technology.2021; 277: 114972. CrossRef
Response of sheep rumen fermentation and microbial communities to feed infected with the endophyte Epichloë gansuensis as evaluated with rumen-simulating technology
Yaling Ma, Hucheng Wang, Chunjie Li
Journal of Microbiology.2021; 59(8): 718. CrossRef
Brisket Disease Is Associated with Lower Volatile Fatty Acid Production and Altered Rumen Microbiome in Holstein Heifers
Naren Gaowa, Kevin Panke-Buisse, Shuxiang Wang, Haibo Wang, Zhijun Cao, Yajing Wang, Kun Yao, Shengli Li
Animals.2020; 10(9): 1712. CrossRef
A Mixed Phytogenic Modulates the Rumen Bacteria Composition and Milk Fatty Acid Profile of Water Buffaloes
Faiz-ul Hassan, Hossam M. Ebeid, Zhenhua Tang, Mengwei Li, Lijuan Peng, Kaiping Peng, Xin Liang, Chengjian Yang
Frontiers in Veterinary Science.2020;[Epub] CrossRef
Comparing the Bacterial Community in the Gastrointestinal Tracts Between Growth-Retarded and Normal Yaks on the Qinghai–Tibetan Plateau
Jian Ma, Yixiao Zhu, Zhisheng Wang, Xiong Yu, Rui Hu, Xueying Wang, Guang Cao, Huawei Zou, Ali Mujtaba Shah, Quanhui Peng, Bai Xue, Lizhi Wang, Suonan Zhao, Xiangying Kong
Frontiers in Microbiology.2020;[Epub] CrossRef
Bacterial communities in the solid, liquid, dorsal, and ventral epithelium fractions of yak (Bos grunniens) rumen
Qingmiao Ren, Huazhe Si, Xiaoting Yan, Chang Liu, Luming Ding, Ruijun Long, Zhipeng Li, Qiang Qiu
MicrobiologyOpen.2020;[Epub] CrossRef
Interactions Between Rumen Microbes, VFAs, and Host Genes Regulate Nutrient Absorption and Epithelial Barrier Function During Cold Season Nutritional Stress in Tibetan Sheep
Xiu Liu, Yuzhu Sha, Renqing Dingkao, Wei Zhang, Weibing Lv, Hong Wei, Hao Shi, Jiang Hu, Jiqing Wang, Shaobin Li, Zhiyun Hao, Yuzhu Luo
Frontiers in Microbiology.2020;[Epub] CrossRef
Yak rumen microbial diversity at different forage growth stages of an alpine meadow on the Qinghai-Tibet Plateau
Li Ma, Shixiao Xu, Hongjin Liu, Tianwei Xu, Linyong Hu, Na Zhao, Xueping Han, Xiaoling Zhang
PeerJ.2019; 7: e7645. CrossRef
Comparing the Microbial Community in Four Stomach of Dairy Cattle, Yellow Cattle and Three Yak Herds in Qinghai-Tibetan Plateau
Jinwei Xin, Zhixin Chai, Chengfu Zhang, Qiang Zhang, Yong Zhu, Hanwen Cao, Jincheng Zhong, Qiumei Ji
Frontiers in Microbiology.2019;[Epub] CrossRef
Dynamic Alterations in Yak Rumen Bacteria Community and Metabolome Characteristics in Response to Feed Type
Chang Liu, Hao Wu, Shujie Liu, Shatuo Chai, Qingxiang Meng, Zhenming Zhou
Frontiers in Microbiology.2019;[Epub] CrossRef

Development and validation of multiplex real-time PCR assays for rapid detection of cytomegalovirus, Epstein-Barr virus, and polyomavirus BK in whole blood from transplant candidates: Kyung-Ah Hwang , Ji Hoon Ahn , Jae-Hwan Nam; J. Microbiol. 2018;56(8):593-599. Published online July 25, 2018; DOI: https://doi.org/10.1007/s12275-018-8273-2

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Abstract

Transplant recipients are more susceptible to bacterial and viral infections. Cytomegalovirus (CMV), Epstein-Barr virus (EBV), and polyomavirus BK (BK) are risk factors for graft dysfunction. All three of them are latent viruses that can cause serious disease in immunocompromised patients. Mainly qualitative PCR tests are required for diagnosis and quantitative monitoring, which are used to follow the response to transplantation. We developed a multiplex real-time PCR (qPCR)
method
to detect these viruses during blood screenings of transplant recipients. We also validated analytical and clinical performance tests using the developed multiplex qPCR. The limit of detection (LOD) was 100, 125, and 183 copies/ml for CMV, EBV, and BK, respectively. These results had high linearity (R2 = 0.997) and reproducibility (CV range, 0.95– 2.38%, 0.52–3.32%, and 0.31–2.45%, respectively). Among 183 samples, we detected 8 samples that were positive for CMV, while only 6 were positive for EBV, and 3 were positive for BK. Therefore, the viral infection prevalence in transplant candidates was 4.40% for CMV, 3.29% for EBV, and 1.64% for BK. This multiplex qPCR method should be used widely for diagnosing and monitoring latent viral infections in transplant recipients.

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CRISPR-Cas-mediated diagnostics
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Transcriptional profiles of laccase genes in the brown rot fungus Postia placenta MAD-R-698: Hongde An , Dongsheng Wei , Tingting Xiao; J. Microbiol. 2015;53(9):606-615. Published online August 1, 2015; DOI: https://doi.org/10.1007/s12275-015-4705-4

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Abstract

One of the laccase isoforms in the brown rot fungus Postia placenta is thought to contribute to the production of hydroxyl radicals, which play an important role in lignocellulose degradation. However, the presence of at least two laccase isoforms in this fungus makes it difficult to understand the details of this mechanism. In this study, we systematically investigated the transcriptional patterns of two laccase genes, Pplcc1 and Pplcc2, by quantitative PCR (qPCR) to better understand the mechanism. The qPCR results showed that neither of the two genes was expressed constitutively throughout growth in liquid culture or during the degradation of a woody substrate. Transcription of Pplcc1 was upregulated under nitrogen depletion and in response to a high concentration of copper in liquid culture, and during the initial colonization of intact aspen wafer. However, it was subject to catabolite repression by a high concentration of glucose. Transcription of Pplcc2 was upregulated by stresses caused by ferulic acid, 2, 6-dimethylbenzoic acid, and ethanol, and under osmotic stress in liquid culture. However, the transcription of Pplcc2 was downregulated upon contact with the woody substrate in solid culture. These results indicate that Pplcc1 and Pplcc2 are differentially regulated in liquid and solid cultures. Pplcc1 seems to play the major role in producing hydroxyl radicals and Pplcc2 in the stress response during the degradation of a woody substrate.

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Isolation, characterization and optimizations of laccase producer from soil: A comprehensive study of application of statistical approach to enhance laccase productivity in Myrothecium verrucaria NFCCI 4363
J.P. Jawale, V.S. Nandre, R.V. Latpate, M.V. Kulkarni, P.J. Doshi
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Kavleen Kaur, Harsimran Sidhu, Neena Capalash, Prince Sharma
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Reference genes for accurate normalization of gene expression in wood-decomposing fungi
Jiwei Zhang, Hugh D. Mitchell, Lye Meng Markillie, Matthew J. Gaffrey, Galya Orr, Jonathan Schilling
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Laccase induction by synthetic dyes in Pycnoporus sanguineus and their possible use for sugar cane bagasse delignification
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Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R
Hongde An, Tingting Xiao, Huan Fan, Dongsheng Wei
Electronic Journal of Biotechnology.2015; 18(6): 451. CrossRef

Functional Analysis of a Subtilisin-like Serine Protease Gene from Biocontrol Fungus Trichoderma harzianum: Haijuan Fan , Zhihua Liu , Rongshu Zhang , Na Wang , Kai Dou , Gulijimila Mijiti , Guiping Diao , Zhiying Wang; J. Microbiol. 2014;52(2):129-138. Published online February 1, 2014; DOI: https://doi.org/10.1007/s12275-014-3308-9

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Abstract

The subtilisin-like serine protease gene ThSS45 has been cloned from Trichoderma harzianum ACCC30371. Its coding region is 1302 bp in length, encoding 433 amino acids, with a predicted protein molecular weight of 44.9 kDa and pI of 5.91. ThSS45 was shown by RT-qPCR analysis to be differentially transcribed in response to eight different treatments. The transcription of ThSS45 was up-regulated when grown in mineral medium, under carbon starvation, and nitrogen starvation, and in the presence of 1% root powder, 1% stem powder, and 1% leaf powder derived from Populus davidiana × P. bolleana (Shanxin poplar) aseptic seedlings. The highest increase in transcription approached 3.5-fold that of the control at 6 h under induction with 1% poplar root powder. The transcription of ThSS45 was also slightly up-regulated by 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid. Moreover, the analyses of coding and promoter regions of ThSS45 homologs indicated that serine protease may be involved in both mycoparasitism and antibiotic secretion. ThSS45 was cloned into the pGEX-4T-2 vector and then expressed in Escherichia coli BL21. The recombinant protein, with an expected molecular weight of approximately 69 kDa, was then purified. When transformant BL21-ss was induced with 1 mM IPTG for 6 h, the purified protease activity reached a peak of 18.25 U/ml at pH 7.0 and 40°C. In antifungal assays the purified protease obviously inhibited the growth of A. alternata mycelia.

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Journal Article

Transcriptional and Biochemical Characterization of Two Azotobacter vinelandii FKBP Family Members: Maria Dimou , Chrysoula Zografou , Anastasia Venieraki , Panagiotis Katinakis; J. Microbiol. 2011;49(4):635-640. Published online September 2, 2011; DOI: https://doi.org/10.1007/s12275-011-0498-2

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Abstract: Peptidyl-prolyl cis/trans isomerases (PPIases, EC: 5.2.1.8), a class of enzymes that catalyse the rate-limiting step of the cis/trans isomerization in protein folding, are divided into three structurally unrelated families: cyclophilins, FK506-binding proteins (FKBPs), and parvulins. Two recombinant FKBPs from the soil nitrogenfixing bacterium Azotobacter vinelandii, designated as AvfkbX and AvfkbB, have been purified and their peptidyl-prolyl cis/trans isomerase activity against Suc-Ala-Xaa-Pro-Phe-pNA synthetic peptides characterised. The substrate specificity of both enzymes is typical for bacterial FKBPs, with Suc-Ala-Phe-Pro-Phe-pNA being the most rapidly catalysed substrate by AvfkbX and Suc-Ala-Leu-Pro-Phe-pNA by AvfkbB. Both FKBPs display chaperone activity as well in the citrate synthase thermal aggregation assay. Furthermore, using real-time RT-qPCR, we demonstrated that both genes were expressed during the exponential growth phase on glucose minimal medium, while their expression declined dramatically during the stationary growth phase as well as when the growth medium was supplied exogenously with ammonium.

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

NOTE] Development of Porphyromonas gingivalis-Specific Quantitative Real-Time PCR Primers Based on the Nucleotide Sequence of rpoB: Soon-Nang Park , Jae-Yoon Park , Joong-Ki Kook; J. Microbiol. 2011;49(2):315-319. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-1028-y

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Abstract: Species-specific quantitative real-time PCR (qPCR) primers were developed for the detection of Porphyromonas gingivalis. These primers, Pg-F/Pg-R, were designed based on the nucleotide sequences of RNA polymerase β-subunit gene (rpoB). Species-specific amplicons were obtained from the tested P. gingivalis strains but not in any of the other strains (46 strains of 46 species). The qPCR primers could detect as little as 4 fg of P. gingivalis chromosomal DNA. These findings suggest that these qPCR primers are suitable for applications in epidemiological studies.

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