Journal of Microbiology

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

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

53 View
0 Download
20 Crossref

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.

Citations

Citations to this article as recorded by

The role of Trichoderma koningii and Trichoderma harzianum in mitigating the combined stresses motivated by Sclerotiniasclerotiorum and salinity in common bean (Phaseolusvulgaris)
Abdelrazek S. Abdelrhim, Nada F. Hemeda, Mai Ali Mwaheb, Maha O.A. Omar, Mona F.A. Dawood
Plant Stress.2024; 11: 100370. CrossRef
Mechanism of oxalate decarboxylase Oxd_S12 from Bacillus velezensis BvZ45-1 in defence against cotton verticillium wilt
Ying Sun, Na Yang, Sirui Li, Fei Chen, Yijing Xie, Canming Tang, Monica Höfte
Journal of Experimental Botany.2024; 75(11): 3500. CrossRef
Purification and Identification of the Nematicidal Activity of S1 Family Trypsin-Like Serine Protease (PRA1) from Trichoderma longibrachiatum T6 Through Prokaryotic Expression and Biological Function Assays
Nan Ma, Hang Lv, Solomon Boamah, Shuwu Zhang, Bingliang Xu
Genes.2024; 15(11): 1437. CrossRef
Genome and transcriptome sequencing of Trichoderma harzianum T4, an important biocontrol fungus of Rhizoctonia solani, reveals genes related to mycoparasitism
Yaping Wang, Jian Wang, Xiaochong Zhu, Wei Wang
Canadian Journal of Microbiology.2024; 70(3): 86. CrossRef
Strain improvement of Trichoderma harzianum for enhanced biocontrol capacity: Strategies and prospects
Ziyang Xiao, Qinqin Zhao, Wei Li, Liwei Gao, Guodong Liu
Frontiers in Microbiology.2023;[Epub] CrossRef
Analysis of the Properties of 44 ABC Transporter Genes from Biocontrol Agent Trichoderma asperellum ACCC30536 and Their Responses to Pathogenic Alternaria alternata Toxin Stress
Hua-Ying Du, Yu-Zhou Zhang, Kuo Liu, Pei-Wen Gu, Shuang Cao, Xiang Gao, Zhi-Ying Wang, Zhi-Hua Liu, Ze-Yang Yu
Current Issues in Molecular Biology.2023; 45(2): 1570. CrossRef
Insights into the ecological generalist lifestyle of Clonostachys fungi through analysis of their predicted secretomes
Edoardo Piombo, Micol Guaschino, Dan Funck Jensen, Magnus Karlsson, Mukesh Dubey
Frontiers in Microbiology.2023;[Epub] CrossRef
The dark septate endophyte Phialocephala sphaeroides suppresses conifer pathogen transcripts and promotes root growth of Norway spruce
Kai Wang, Zilan Wen, Fred O Asiegbu, Malin Elfstrand
Tree Physiology.2022; 42(12): 2627. CrossRef
Extracellular proteins of Trichoderma and their role in plant health
Anu Sharma, Richa Salwan, Vivek Sharma
South African Journal of Botany.2022; 147: 359. CrossRef
Predicted Input of Uncultured Fungal Symbionts to a Lichen Symbiosis from Metagenome-Assembled Genomes
Gulnara Tagirdzhanova, Paul Saary, Jeffrey P Tingley, David Díaz-Escandón, D Wade Abbott, Robert D Finn, Toby Spribille, Jason Stajich
Genome Biology and Evolution.2021;[Epub] CrossRef
Production of tailor-made enzymes to facilitate lipid extraction from the oleaginous yeast Schwanniomyces occidentalis
Ruud Heshof, Bram Visscher, Eric van de Zilver, Rick van de Vondervoort, Femke van Keulen, Roy J. B. M. Delahaije, Richèle D. Wind
AMB Express.2020;[Epub] CrossRef
An approach to select Lactobacillus isolates as protective cultures for food fermentations
Raffael C. Inglin, Alessia I. Delbrück, Benjamin Fässler, Katharina E. Siebenmann, Christophe Lacroix, Marc J. A. Stevens, Leo Meile
Journal of Food Safety.2018;[Epub] CrossRef
Biocontrol activity of recombinant aspartic protease from Trichoderma harzianum against pathogenic fungi
Jun-Jin Deng, Wei-Qian Huang, Zhi-Wei Li, De-Lin Lu, Yuanyuan Zhang, Xiao-chun Luo
Enzyme and Microbial Technology.2018; 112: 35. CrossRef
Functional analysis of eliciting plant response protein Epl1-Tas from Trichoderma asperellum ACCC30536
Wenjing Yu, Gulijimila Mijiti, Ying Huang, Haijuan Fan, Yucheng Wang, Zhihua Liu
Scientific Reports.2018;[Epub] CrossRef
Comparative evolutionary histories of fungal proteases reveal gene gains in the mycoparasitic and nematode-parasitic fungus Clonostachys rosea
Mudassir Iqbal, Mukesh Dubey, Mikael Gudmundsson, Maria Viketoft, Dan Funck Jensen, Magnus Karlsson
BMC Evolutionary Biology.2018;[Epub] CrossRef
Expression analysis on mycoparasitism related genes during antagonism of Trichoderma with Colletotrichum falcatum causing red rot in sugarcane
Elangovan Elamathi, Palaniyandi Malathi, Rasappa Viswanathan, Amalraj Ramesh Sundar
Journal of Plant Biochemistry and Biotechnology.2018; 27(3): 351. CrossRef
Subtilisin-like serine protease gene TghSS42 from Trichoderma ghanense ACCC 30153 was successfully expressed in Escherichia coli and recombinant protease rTghSS42 exhibited antifungal ability to five phytopathogens
HUIFANG ZHANG, NA WANG, YUCHENG WANG, JINJIE WANG, HONG ZHENG, ZHIHUA LIU
Biocontrol Science.2017; 22(3): 145. CrossRef
A novel organic solvent- and detergent-stable serine alkaline protease from Trametes cingulata strain CTM10101
Maroua Omrane Benmrad, Emna Moujehed, Mouna Ben Elhoul, Nadia Zaraî Jaouadi, Sondes Mechri, Hatem Rekik, Sidali Kourdali, Mohamed El Hattab, Abdelmalek Badis, Sami Sayadi, Samir Bejar, Bassem Jaouadi
International Journal of Biological Macromolecules.2016; 91: 961. CrossRef
Differential Response of Extracellular Proteases of Trichoderma Harzianum Against Fungal Phytopathogens
Vivek Sharma, Richa Salwan, Prem N. Sharma
Current Microbiology.2016; 73(3): 419. CrossRef
Fungal proteins and genes associated with biocontrol mechanisms of soil-borne pathogens: a review
Yohann Daguerre, Katarzyna Siegel, Véronique Edel-Hermann, Christian Steinberg
Fungal Biology Reviews.2014; 28(4): 97. CrossRef

Characterization, Cloning, and Heterologous Expression of a Subtilisin-Like Serine Protease Gene VlPr1 from Verticillium lecanii: Gang Yu , Jin-Liang Liu , Li-Qin Xie , Xue-Liang Wang , Shi-Hong Zhang , Hong-Yu Pan; J. Microbiol. 2012;50(6):939-946. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2199-x

37 View
0 Download
12 Scopus

Abstract: The entomopathogenic fungus Verticillium lecanii is a wellknown biocontrol agent. V. lecanii produces subtilisin-like serine protease (Pr1), which is important in the biological control activity of some insect pests by degrading insect cuticles. In this study, a subtilisin-like serine protease gene VlPr1 was cloned from the fungus and the VlPr1 protein was expressed in Escherichia coli. The VlPr1 gene contains an open reading frame (ORF) interrupted by three short introns, and encodes a protein of 379 amino acids. Protein sequence analysis revealed high homology with subtilisin serine proteases. The molecular mass of the protease was 38 kDa, and the serine protease exhibited its maximal activity at 40°C and pH 9.0. Protease activity was also affected by Mg2+ and Ca2+ concentration. The protease showed inhibitory activity against several plant pathogens, especially towards Fusarium moniliforme.

Journal Article

Cloning, Expression, and Characterization of Serine Protease from Thermophilic Fungus Thermoascus aurantiacus var. levisporus: An-Na Li , Chen Xie , Jie Zhang , Jia Zhang , Duo-Chuan Li; J. Microbiol. 2011;49(1):121-129. Published online March 3, 2011; DOI: https://doi.org/10.1007/s12275-011-9355-6

27 View
0 Download
17 Scopus

Abstract: The serine protease gene from a thermophilic fungus Thermoascus aurantiacus var. levisporus, was cloned, sequenced, and expressed in Pichia pastoris and the recombinant protein was characterized. The full-length cDNA of 2,592 bp contains an ORF of 1,482 bp encoding 494 amino acids. Sequence analysis of the deduced amino acid sequence revealed high homology with subtilisin serine proteases. The putative enzyme contained catalytic domain with active sites formed by three residues of Asp183, His215, and Ser384. The molecular mass of the recombinant enzyme was estimated to be 59.1 kDa after overexpression in P. pastoris. The activity of recombinant protein was 115.58 U/mg. The protease exhibited its maximal activity at 50°C and pH 8.0 and kept thermostable at 60°C, and retained 60% activity after 60 min at 70°C. The protease activity was found to be inhibited by PMSF, but not by DTT or EDTA. The enzyme has broad substrate specificity such as gelatin, casein and pure milk, and exhibiting highest activity towards casein.

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

Taxonomic Revision of the Nematode-Trapping Fungus Arthrobotrys multisecundaria: Juan Li , Jinkui Yang , Lianming Liang , Ke-Qin Zhang; J. Microbiol. 2008;46(5):513-518. Published online October 31, 2008; DOI: https://doi.org/10.1007/s12275-007-0115-6

41 View
0 Download
2 Scopus

Abstract: The gene encoding an extracellular serine protease was cloned from Arthrobotrys multisecundaria using degenerate primers. The gene was highly similar (99.26%) to protease Mlx from Monacrosporium microscaphoides. To clarify the taxonomic relationship between these species, genes encoding the internal transcribed spacer (ITS) and β-tubulin were also cloned and sequenced from A. multisecundaria and M. microscaphoides, respectively. Homologous analysis of the nuclear (ITS) and protein (β-tubulin) encoding genes showed that the two species of nematode-trapping fungi also shared extensive identity (99.82 and 99.63%, respectively), although they exhibited obvious differences in secondary conidia morphology. Accordingly, a taxonomic revision is recommended, with A. multisecundaria being revised as A. microscaphoides var. multisecundaria. In addition, the identified mutation may better facilitate the study of the sporulation of nematode-trapping fungi.

A Fibrinolytic Enzyme from the Medicinal Mushroom Cordyceps militaris: Jae-Sung Kim , Kumar Sapkota , Se-Eun Park , Bong-Suk Choi , Seung Kim , Nguyen Thi Hiep , Chun-Sung Kim , Han-Seok Choi , Myung-Kon Kim , Hong-Sung Chun , Yeal Park , Sung-Jun Kim; J. Microbiol. 2006;44(6):622-631.; DOI: https://doi.org/2465 [pii]

39 View
0 Download

Abstract: In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9%. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and 37°C, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin α-chain followed by the γ-γ chains. It also hydrolyzed the β-chain, but more slowly. The Aα, Bβ, and γ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by Cu2+ and Co2+, but enhanced by the additions of Ca2+ and Mg2+ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it’s a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

Multicatalytic Alkaline Serine Protease from the Psychrotrophic Bacillus amyloliquefaciens S94: Eui-Sun Son , Jong-Il Kim; J. Microbiol. 2003;41(1):58-62.

35 View
0 Download

Abstract: An extracellular protease of Bacillus amyloliquefaciens S94 was purified to apparent homogeneity. The enzyme activity was strongly inhibited by general inhibitor for serine protease, PMSF, suggesting that the enzyme is a serine protease. The purified enzyme activity was inhibited by leucine peptidase inhibitor, bestatin, suggesting that the enzyme is a leucine endopeptidase. The maximum proteolytic activity against different protein substrates occurred at pH 10, 45℃ (protein substrate) and pH 8, 45℃ (synthetic substrate). The purified enzyme was specific in that it readily hydrolyzed substrates with Leu or Lys residues at P1 site. The protease had characteristics of a cold-adapted protein, which was more active for the hydrolysis of synthetic substrate in the range of 15℃ to 45℃, specially at low temperature.

A Recombinant Human [alpha]_1-Antitrypsin Variant, M_malton, Undergoes a Spontaneous Conformational Conversion into a Latent Form: Chan-Hun Jung , Hana Im; J. Microbiol. 2003;41(4):335-339.

34 View
0 Download

Abstract: Many genetic variants of [alpha]_1-antitrypsin have been associated with early onset emphysema and liver cirrhosis. However, the detailed structural basis of pathogenic [alpha]_1-antitrypsin molecules is rarely known. Here we found that a recombinant M_malton variant (Phe52-deleted) lost inhibitory activity by spontaneous conformational conversion into a more stable, inactive form under physiological conditions. Biochemical and spectroscopic data suggested that the variant converts into a reactive center loop-inserted conformation, resembling the latent form of plasminogen activator inhibitor-1.

Streptomyces griseus Trypsin (SGT) Has Gelatinase Activity and Its Proteolytic Activity Is Enhanced by Manganese: Won-Jae Chi , Yoon-Hee Kim , Jong-Hee Kim , Dae-Kyung Kang , Sang-Soon Kang , Joo-Won Suh , Soon-Kwang Hong; J. Microbiol. 2003;41(4):289-294.

34 View
0 Download

Abstract: Gelatinase is a proteolytic enzyme that hydrolyzes gelatin. Gelatinolytic activity was detected from culture broths of Streptomyces griseus IFO13350 and HH1 by paper disc assays on 0.5% agar plates containing 1% gelatin. The concentrated extracellular protein from the S. griseus was analyzed by SDS polyacrylamide gel, and two proteins, with molecular weights of 30 and 28 kDa, respectively, were identified to have gelatinase activity by gelatin zymography. The protein with a molecular weight of 28 kDa was confirmed to be S. griseus trypsin (SGT). The effects of metal ions and metal chelators on the protease activity of the SGT were studied. Of the metal ions tested, only manganese was found to enhance the protease activity, 2.6 times, however, Co_2^+, Cu_2^+, and Zn_2^+, and metal chelators, such as EDTA and EGTA, inhibited the SGT activity. When the protease activity of the SGT was measured at various pHs, in the presence of 5 mM MnCl_2, its highest activity was at pH 11.0, whereas only 60% of the maximum activity was observed between pHs 4.0 and pH 6.0, and almost 80% activity between pHs 7.0 to pH 10.0. The protease activity was measured at various temperatures in the presence of 5 mM MnCl_2. The SGT was found to be stable up to 60℃ for 30 min, while only 16% of the enzyme activity remained at 60℃, and at 80℃ almost all the activity was lost. The optimal temperature for the protease activity was 50℃.

First
Prev
Page of 1
Next
Last

Search