Abstract:
:This study shows that sequential introduction of drug resistance mutations substantially increased enzyme production in Paenibacillus agaridevorans The triple mutant YT478 (rsmG Gln225→stop codon, rpsL K56R, and rpoB R485H), generated by screening for resistance to streptomycin and rifampin, expressed a 1,100-fold-larger amount of the extracellular enzyme cycloisomaltooligosaccharide glucanotransferase (CITase) than the wild-type strain. These mutants were characterized by higher intracellular S-adenosylmethionine concentrations during exponential phase and enhanced protein synthesis activity during stationary phase. Surprisingly, the maximal expression of CITase mRNA was similar in the wild-type and triple mutant strains, but the mutant showed greater CITase mRNA expression throughout the growth curve, resulting in enzyme overproduction. A metabolome analysis showed that the triple mutant YT478 had higher levels of nucleic acids and glycolysis metabolites than the wild type, indicating that YT478 mutant cells were activated. The production of CITase by the triple mutant was further enhanced by introducing a mutation conferring resistance to the rare earth element, scandium. This combined drug resistance mutation method also effectively enhanced the production of amylases, proteases, and agarases by P. agaridevorans and Streptomyces coelicolor This method also activated the silent or weak expression of the P. agaridevorans CITase gene, as shown by comparisons of the CITase gene loci of P. agaridevorans T-3040 and another cycloisomaltooligosaccharide-producing bacterium, Paenibacillus sp. strain 598K. The simplicity and wide applicability of this method should facilitate not only industrial enzyme production but also the identification of dormant enzymes by activating the expression of silent or weakly expressed genes.IMPORTANCE Enzyme use has become more widespread in industry. This study evaluated the molecular basis and effectiveness of ribosome engineering in markedly enhancing enzyme production (>1,000-fold). This method, due to its simplicity, wide applicability, and scalability for large-scale production, should facilitate not only industrial enzyme production but also the identification of novel enzymes, because microorganisms contain many silent or weakly expressed genes which encode novel antibiotics or enzymes. Furthermore, this study provides a new mechanism for strain improvement, with a consistent rather than transient high expression of the key gene(s) involved in enzyme production.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Funane K,Tanaka Y,Hosaka T,Murakami K,Miyazaki T,Shiwa Y,Gibu S,Inaoka T,Kasahara K,Fujita N,Yoshikawa H,Hiraga Y,Ochi Kdoi
10.1128/JB.00188-18subject
Has Abstractpub_date
2018-08-10 00:00:00issue
17eissn
0021-9193issn
1098-5530pii
JB.00188-18journal_volume
200pub_type
杂志文章abstract::Growth and sporulation of a Bacillus subtilis mutant deficient in branched fatty acid synthesis (gene symbol bfmB) were examined. The mutant, which produces an acyl-coenzyme A:acyl carrier protein transacylase with reduced affinity for branched fatty acid primers, could grow in media containing any one of a wide range...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.148.2.480-486.1981
更新日期:1981-11-01 00:00:00
abstract::The Lyme disease spirochete Borrelia burgdorferi is unique among bacteria in its large number of lipoproteins that are encoded by a small, exceptionally fragmented, and predominantly linear genome. Peripherally anchored in either the inner or outer membrane and facing either the periplasm or the external environment, ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00658-16
更新日期:2017-02-28 00:00:00
abstract::For maximal turbidity and dissimilatory activity, three strains of pediococci required the addition of Mn(2+) to Rogosa basal medium. Twenty-two strains of enterococci proved indifferent to this supplement. ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.112.1.627-628.1972
更新日期:1972-10-01 00:00:00
abstract::Growth of Streptococcus faecalis in the presence of l-malate resulted in the induction of a "malic enzyme" [l-malate:nicotinamide adenine dinucleotide (NAD) oxidoreductase (decarboxylating), E.C. 1.1.1.39]. Synthesis of the malic enzyme did not appear to be subject to catabolite repression by intermediate products of ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.98.2.705-711.1969
更新日期:1969-05-01 00:00:00
abstract::The Ssa subfamily of Hsp70 molecular chaperones in the budding yeast Saccharomyces cerevisiae has four members, encoded by SSA1, SSA2, SSA3, and SSA4. Deletion of the two constitutively expressed genes, SSA1 and SSA2, results in cells which are slow growing and temperature sensitive. In this study, we demonstrate that...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.24.6484-6492.1998
更新日期:1998-12-01 00:00:00
abstract::Germination of Bacillus spores is triggered by the binding of specific nutrients to germinant receptors (GRs) located in the spore's inner membrane. The GRs typically consist of A, B, and C subunits, encoded by tricistronic ger operons. The Bacillus licheniformis genome contains the gerA family operons gerA, ynd, and ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00451-19
更新日期:2019-10-04 00:00:00
abstract::A new type of temperature-sensitive deoxyribonucleic acid (DNA) synthesis mutant, which can divide without a completion of DNA replication, was isolated from a thymidine-requiring Escherichia coli strain by means of photo-bromouracil selection after nitrosoguanidine mutagenesis. In this mutant, in spite of the fact th...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.99.3.842-850.1969
更新日期:1969-09-01 00:00:00
abstract::Pili isolated from Pseudomonas aeruginosa PAK were solubilized in the detergent octyl-glucoside. Subsequent removal of the detergent by dialysis resulted in the formation of short rods of about 9 nm in diameter and various lengths. The aggregation process was followed by ultracentrifugation, viscometry, and electron m...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.151.3.1508-1513.1982
更新日期:1982-09-01 00:00:00
abstract::Flagellum formation by ubiquinone- and menaquinone-deficient mutant strains of Escherichia coli K-12 was studied under both aerobic and anaerobic growth conditions. Ubiquinone was found to be obligatory for aerobic flagellum formation but could be replaced by menaquinone for anaerobic flagellum formation. A mutant dev...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.132.3.1034-1035.1977
更新日期:1977-12-01 00:00:00
abstract::Three evolutionarily distinct families of replicative DNA polymerases, designated polymerase B (Pol B), Pol C, and Pol D, have been identified. Members of the Pol B family are present in all three domains of life, whereas Pol C exists only in Bacteria and Pol D exists only in Archaea. Pol B enzymes replicate eukaryoti...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.02037-12
更新日期:2013-05-01 00:00:00
abstract::Flagellate bacteria such as Escherichia coli and Salmonella enterica serovar Typhimurium typically express 5 to 12 flagellar filaments over their cell surface that rotate in clockwise (CW) and counterclockwise directions. These bacteria modulate their swimming direction towards favorable environments by biasing the di...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.187.5.1695-1701.2005
更新日期:2005-03-01 00:00:00
abstract::The RpoS sigma factor (σ(S)) is the master regulator of the bacterial response to a variety of stresses. Mutants in rpoS arise in bacterial populations in the absence of stress, probably as a consequence of a subtle balance between self-preservation and nutritional competence. We characterized here one natural rpoS mu...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00801-10
更新日期:2010-12-01 00:00:00
abstract::Daniel, John W. (University of Wisconsin, Madison), Karlee L. Babcock, Alice H. Sievert, and Harold P. Rusch. Organic requirements and synthetic media for growth of the myxomycete Physarum polycephalum. J. Bacteriol. 86:324-331. 1963.-An isolate of Physarum polycephalum was maintained in submersed pure culture in a co...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.86.2.324-331.1963
更新日期:1963-08-01 00:00:00
abstract::Here we describe that the Helicobacter pylori sensor kinase produced by HP1364 and the response regulator produced by HP1365 and designated CrdS and CrdR, respectively, are both required for transcriptional induction of the H. pylori copper resistance determinant CrdA by copper ions. CrdRS-deficient mutants lacked cop...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.187.13.4683-4688.2005
更新日期:2005-07-01 00:00:00
abstract::A preparation of pure protoplasts of Geotrichum candidum became osmotically stable and colonies developed when the protoplasts were embedded in stabilizing thin-layer-agar and incubated with stabilizing basal medium. When growing protoplasts were exposed to distilled water and then reincubated with basal medium, the p...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.98.1.256-263.1969
更新日期:1969-04-01 00:00:00
abstract::We studied the roles of Streptococcus thermophilus phosphogalactosyltransferase (EpsE) (the priming enzyme), tyrosine kinase (EpsD), phosphatase (EpsB), and a membrane-associated protein with no known biochemical function (EpsC) in exopolysaccharide (EPS) synthesis. These proteins are well-conserved among bacteria and...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01122-06
更新日期:2007-02-01 00:00:00
abstract::Forty years ago, Coulter and Talalay (A. W. Coulter and P. Talalay, J. Biol. Chem. 243:3238-3247, 1968) established the oxygenase-dependent pathway for the degradation of testosterone by aerobes. The oxic testosterone catabolic pathway involves several oxygen-dependent reactions and is not available for anaerobes. Sin...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00331-11
更新日期:2011-09-01 00:00:00
abstract::The distribution of a pulse of teichoic acid-specific radiolabel between wall and membrane teichoic acids in pneumococci was constant over a subsequent chase period, suggesting that wall and membrane teichoic acids are biosynthesized simultaneously and independently. ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.122.1.335-337.1975
更新日期:1975-04-01 00:00:00
abstract::Kobayashi, George S. (Tulane University, New Orleans, La.), Lorraine Friedman, and Judith F. Kofroth. Some cytological and pathogenic properties of spheroplasts of Candida albicans. J. Bacteriol. 88:795-801. 1964.-Spheroplasts of Candida albicans were prepared by use of an enzymatic mixture from the digestive tract of...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.88.3.795-801.1964
更新日期:1964-09-01 00:00:00
abstract::The so-called symbiotic region of the Bradyrhizobium japonicum chromosome (C. Kündig, H. Hennecke, and M. Göttfert, J. Bacteriol. 175:613-622, 1993) was screened for the presence of genes controlled by the nitrogen fixation regulatory protein NifA. Southern blots of restriction enzyme-digested cosmids that represent a...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.6.1472-1480.2000
更新日期:2000-03-01 00:00:00
abstract::Clostridium difficile, a proteolytic Gram-positive anaerobe, has emerged as a significant nosocomial pathogen. Stickland fermentation reactions are thought to be important for growth of C. difficile and appear to influence toxin production. In Stickland reactions, pairs of amino acids donate and accept electrons, gene...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01492-12
更新日期:2013-02-01 00:00:00
abstract::This study describes a novel approach to identify unique genomic DNA sequences from the unsequenced strain C. jejuni ATCC 43431 by comparison with the sequenced strain C. jejuni NCTC 11168. A shotgun DNA microarray was constructed by arraying 9,600 individual DNA fragments from a C. jejuni ATCC 43431 genomic library o...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.186.14.4781-4795.2004
更新日期:2004-07-01 00:00:00
abstract::Escherichia coli W3110 contains four types of IS1 elements in the chromosome. Using an insertion element entrapping system, we collected 116 IS1 plasmid insertion mutants, which resulted from a minimum of 26 independent IS1 insertion events. All of them had insertions of IS1 of the IS1A (IS1E and IS1G) type. Inspectio...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.19.5279-5283.1998
更新日期:1998-10-01 00:00:00
abstract::PhoB is the response regulator of the Pho regulon. It is composed of two distinct domains, an N-terminal receiver domain and a C-terminal output domain that binds DNA and interacts with sigma(70) to activate transcription of the Pho regulon. Phosphorylation of the receiver domain is required for activation of the prot...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.23.6592-6597.2000
更新日期:2000-12-01 00:00:00
abstract::The bacteriophage T4 MotA protein is a transcriptional activator of T4-modified host RNA polymerase and is required for activation of the middle class of T4 promoters. MotA alone binds to the -30 region of T4 middle promoters, a region that contains the MotA box consensus sequence [(t/a)(t/a)TGCTT(t/c)A]. We report th...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.178.21.6133-6139.1996
更新日期:1996-11-01 00:00:00
abstract::The AOX1 gene, which encodes an alternative oxidase, was isolated from the genomic DNA library of Candida albicans. The gene encodes a polypeptide consisting of 379 amino acids with a calculated molecular mass of 43,975 Da. The aox1/aox1 mutant strain did not show cyanide-resistant respiration under normal conditions ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.181.13.4098-4102.1999
更新日期:1999-07-01 00:00:00
abstract::The protein motive force of metabolizing Bacillus subtilis cells was only slightly affected by changes in the external pH between 5 and 8, although the electrical component and the chemical component of the proton motive force contributed differently at different external pH. The electrical component of the proton mot...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.144.3.891-897.1980
更新日期:1980-12-01 00:00:00
abstract::We report the draft genome sequence of Lactobacillus salivarius SMXD51, isolated from the cecum of healthy chickens showing an activity against Campylobacter--the food-borne pathogen that is the most common cause of gastroenteritis in the European Union (EU)--and potentially interesting features for a probiotic strain...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00344-12
更新日期:2012-06-01 00:00:00
abstract::A spontaneously arising regulatory mutant of the gluconate system in Escherichia coli was isolated. This mutant became constitutive, probably in one step, for gluconate high-affinity transport, gluconokinase, and gluconate-6-P dehydrase. The mutation involved (gntR18) is cotransducible with asd. Pseudorevertants, deri...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.114.2.469-473.1973
更新日期:1973-05-01 00:00:00
abstract::The activity of the ATP:GTP 3'-pyrophosphotransferase (guanosine pentaphosphate synthetase I [GPSI]) from Streptomyces antibioticus is stimulated maximally by methanol at 20% (vol/vol) in assay mixtures. Although the enzyme is not activated by ribosomes, its activity is stimulated by tRNA (uncharged or charged) and by...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.176.5.1482-1487.1994
更新日期:1994-03-01 00:00:00