Abstract:
:Genome sequencing of the thermophilic archaeon Pyrococcus horikoshii OT3 revealed a gene which had high sequence similarity to the gene encoding the carboxypeptidase of Sulfolobus solfataricus and also to that encoding the aminoacylase from Bacillus stearothermophilus. The gene from P. horikoshii comprises an open reading frame of 1,164 bp with an ATG initiation codon and a TGA termination codon, encoding a 43,058-Da protein of 387 amino acid residues. However, some of the proposed active-site residues for carboxypeptidase were not found in this gene. The gene was overexpressed in Escherichia coli with the pET vector system, and the expressed enzyme had high hydrolytic activity for both carboxypeptidase and aminoacylase at high temperatures. The enzyme was stable at 90 degrees C, with the highest activity above 95 degrees C. The enzyme contained one bound zinc ion per one molecule that was essential for the activity. The results of site-directed mutagenesis of Glu367, which corresponds to the essential Glu270 in bovine carboxypeptidase A and the essential Glu in other known carboxypeptidases, revealed that Glu367 was not essential for this enzyme. The results of chemical modification of the SH group and site-directed mutagenesis of Cys102 indicated that Cys102 was located at the active site and was related to the activity. From these findings, it was proven that this enzyme is a hyperthermostable, bifunctional, new zinc-dependent metalloenzyme which is structurally similar to carboxypeptidase but whose hydrolytic mechanism is similar to that of aminoacylase. Some characteristics of this enzyme suggested that carboxypeptidase and aminoacylase might have evolved from a common origin.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Ishikawa K,Ishida H,Matsui I,Kawarabayasi Y,Kikuchi Hdoi
10.1128/AEM.67.2.673-679.2001keywords:
subject
Has Abstractpub_date
2001-02-01 00:00:00pages
673-9issue
2eissn
0099-2240issn
1098-5336journal_volume
67pub_type
杂志文章abstract::Streptomyces exfoliatus SMF19 produced two types of extracellular beta-lactamase-inhibiting proteins, 48 (beta LIP-I) and 33 (beta LIP-II) kDa. The inhibition mode of beta LIP-I on Bacto Penase and benzylpenicillin as substrates was uncompetitive (i.e., both Km and Vmax were changed), while that of beta LIP-II was non...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.60.3.1029-1032.1994
更新日期:1994-03-01 00:00:00
abstract::Degradation of cotton cellulose by Trichoderma reesei endoglucanase I (EGI) and cellobiohydrolase II (CBHII) was investigated by analyzing the insoluble cellulose fragments remaining after enzymatic hydrolysis. Changes in the molecular-size distribution of cellulose after attack by EGI, alone and in combination with C...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.62.8.2883-2887.1996
更新日期:1996-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.31.5.784-786.1976
更新日期:1976-05-01 00:00:00
abstract::Media used to determine the MICs of heavy metals for bacteria are unreliable because organic components in the media bind or chelate most of the metal being studied. To define specific metal activity in media and to maintain metal activity at a constant level, HEPES-MES [N-2-hydroxyethylpiperazine-N' -2-ethanesulfonic...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.55.8.2101-2104.1989
更新日期:1989-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.49.1.232-233.1985
更新日期:1985-01-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.65.5.1991-1997.1999
更新日期:1999-05-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.51.5.1056-1062.1986
更新日期:1986-05-01 00:00:00
abstract::A fragment of about 500 base pairs of the beta-hemolysin gene from Listeria monocytogenes was used to screen different bacterial strains by DNA colony hybridization. The cells in the colonies were lysed by microwaves in the presence of sodium hydroxide. Of 52 different strains of Listeria species screened, only the DN...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.53.9.2256-2259.1987
更新日期:1987-09-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.50.4.1110-1111.1985
更新日期:1985-10-01 00:00:00
abstract::Lysostaphin represents a promising therapeutic agent for the treatment of staphylococcal infections, in particular those of methicillin-resistant Staphylococcus aureus (MRSA). However, conventional expression systems for the enzyme suffer from various limitations, and there remains a need for an efficient and cost-eff...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.03914-13
更新日期:2014-05-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.57.7.2101-2103.1991
更新日期:1991-07-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.71.11.7376-7388.2005
更新日期:2005-11-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.67.3.1396-1399.2001
更新日期:2001-03-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.71.10.5719-5727.2005
更新日期:2005-10-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.36.5.688-692.1978
更新日期:1978-11-01 00:00:00
abstract::Pure cultures of Escherichia coli and Streptococcus faecalis and environmental water samples were examined for the possibility that pollution involving organotin compounds could decrease the values for indicator organisms when standard methods were applied to the analysis of water samples. (CH3)2SnCl2 and (CH3)3SnCl d...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.52.3.562-566.1986
更新日期:1986-09-01 00:00:00
abstract::This study was initiated to determine if reduced sulfur contained in coal gasifier ash and slag was available for microbial and chemical oxidation because eventual large-quantity landfill disposal of these solid wastes is expected. Continuous application of distilled water to a column containing a high-sulfur-content ...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.45.3.743-747.1983
更新日期:1983-03-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01489-06
更新日期:2007-04-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2013-07-01 00:00:00
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journal_title:Applied and environmental microbiology
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更新日期:2016-07-29 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.59.5.1602-1606.1993
更新日期:1993-05-01 00:00:00
abstract::A method was developed to deposit Bacillus subtilis spores via aerosolization onto various surface materials for biological agent decontamination and detection studies. This new method uses an apparatus coupled with a metered dose inhaler to reproducibly deposit spores onto various surfaces. A metered dose inhaler was...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02237-10
更新日期:2011-03-01 00:00:00
abstract::Several recent outbreaks of toxoplasmosis were related to drinking water. We propose a strategy for Toxoplasma oocyst detection as part of an approach to detecting multiple waterborne parasites, including Giardia and Cryptosporidium spp., by the U.S. Environmental Protection Agency method with the same sample. Water s...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.70.7.4035-4039.2004
更新日期:2004-07-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.00322-14
更新日期:2014-07-01 00:00:00
abstract::Trichoderma reesei can produce up to 100 g/liter of extracellular proteins. The major and industrially relevant products are cellobiohydrolase I (CBHI) and the hemicellulase XYNI. The genes encoding both enzymes are transcriptionally activated by the regulatory protein Xyr1. The first 850 nucleotides of the cbh1 promo...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01742-17
更新日期:2017-12-15 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.55.6.1375-1379.1989
更新日期:1989-06-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01293-06
更新日期:2007-01-01 00:00:00
abstract::Environmental bacteria are exposed to a myriad of biotic interactions that influence their function and survival. The grazing activity of protozoan predators significantly impacts the dynamics, diversification, and evolution of bacterial communities in soil ecosystems. To evade protozoan predation, bacteria employ var...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01272-09
更新日期:2009-11-01 00:00:00
abstract::IncQ marker plasmids were previously constructed to enable the analysis of the survival of populations of Pseudomonas putida released into lake water (C. Winstanley, J. A. W. Morgan, R. W. Pickup, J. G. Jones, and J. R. Saunders, Appl. Environ. Microbiol. 55:771-777, 1989). We constructed equivalent IncP plasmids, pLV...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.57.7.1905-1913.1991
更新日期:1991-07-01 00:00:00
abstract::Cyanobacteria can synthesize alkanes and alkenes, which are considered to be infrastructure-compatible biofuels. In terms of physiological function, cyanobacterial hydrocarbons are thought to be essential for membrane flexibility for cell division, size, and growth. The genetic basis for the biosynthesis of terminal o...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.00425-18
更新日期:2018-06-18 00:00:00