• Journal of Microbiology and Biotechnology
• /
• 제10권6호
• /
• pp.829-835
• /
• 2000

The ptsH and ptsI genes of Lactococus lactis subsp. lactis ATCC 7962 (L. lactis 7962), encoding the general proteins of phosphotransferase system (PTS) components, HPr and enzyme I, respectively, were cloned and characterized. A 1.3 kb PCR product was obtained using a primer set that was hybridized to the internal region of the L. lactis 7962 pts HI genes and then subcloned into a low-copy number vector, pACYC184. The 5' upstream and 3' downstream region from the 1.3 kb fragment were subsequently clone using the chromosome walking method. The complete ptsHI operon was constructed and the nucleotide sequences determined. Two ORFs corresponding to HPr (88 amino acids) and enzyme I (575 amino acids) were located. The ptsHI genes of L. lactis 7962 showed a very high homology (84-90%) with those genes from other Gram-positive bacteria. A primer extension analysis showed that the transcription started at either one of two adjacent bases upstream of the start codon. Using a Northern analysis, two transcripts were detected; the first, a 0.3 kb transcript corresponding to ptsH and the second, a 2 kb transcript corresponding to ptsH and ptsI. The transcription level of ptsH was higher than that of ptsI. The concentration of the ptsH transcript in cells grown on glucose was similar to that in cells grown on lactose, yet higher than that in cells grown on galactose. The ptsI transcript was scarcely detected in cell grown on lactose or galactose. The ptsI transcript was scarcely detected in cells grown on lactose or galactose. The results of a sequence analysis and Northern blot confirmed that the ptsH and ptsI genes of L. lactis 7962 were arranged in an operon like other known ptsHI genes and the expression of the ptsHI genes was regulated at the transcriptional level in response to the carbon source.

• Journal of Microbiology and Biotechnology
• /
• 제23권5호
• /
• pp.656-660
• /
• 2013

Pseudomonas fluorescens strain CL0145A was discovered at the New York State Museum Field Research Laboratory as an effective agent against the environmentally destructive zebra mussel, which has contaminated US waters. Dried cells of the microbe are being commercialized as an environmentally friendly solution to the problem. We found that antibiotic activity against the Gram-positive bacterium Bacillus subtilis is produced and excreted by this strain. We have carried out studies to optimize production of the antibiotic. Studies were begun in a complex corn meal medium. Activity was found in both cells and culture supernates and was maximal after one day of fermentation. Static fermentation conditions were found to be superior to shaken culture. Production of extracellular antibiotic in complex medium was found to be dependent on the content of sucrose and enzyme-hydrolyzed casein. Indeed, production was greater in sucrose plus enzyme-hydrolyzed casein than in the complex medium. Of a large number of carbon sources studied as improvements over sucrose, the best was glycerol. An examination of nitrogen sources showed that production was improved by replacement of enzyme-hydrolyzed casein with soy hydrolysates. Production in the simple glycerol-Hy-Soy medium was not improved by addition of an inorganic salt mixture or by complex nitrogen sources, with the exception of malt extract. In an attempt to keep the medium more defined, we studied the effect of amino acids and vitamins as replacements for malt extract. Of 21 amino acids and 7 vitamins, we found tryptophan, glutamine, biotin, and riboflavin to be stimulatory. The final medium contained glycerol, Hy-Soy, tryptophan, glutamine, biotin, and riboflavin.

• Journal of Microbiology and Biotechnology
• /
• 제29권3호
• /
• pp.373-381
• /
• 2019

Site-directed mutagenesis was employed to generate five different triple point mutations in the double mutant (C295A/I86A) of Thermoanaerobacter ethanolicus alcohol dehydrogenase (TeSADH) by computer-aided modeling with the aim of widening the small alkyl-binding pocket. TeSADH engineering enables the enzyme to accept sterically hindered substrates that could not be accepted by the wild-type enzyme. The underline in the mutations highlights the additional point mutation on the double mutant TeSADH introduced in this work. The catalytic efficiency ($k_{cat}/K_M$) of the ${\underline{M151A}}$/C295A/I86A triple TeSADH mutant for acetophenone increased about 4.8-fold higher than that of the double mutant. A 2.4-fold increase in conversion of 3'-methylacetophenone to (R)-1-(3-methylphenyl)-ethanol with a yield of 87% was obtained by using ${\underline{V115A}}$/C295A/I86A mutant in asymmetric reduction. The ${\underline{A85G}}$/C295A/I86A mutant also produced (R)-1-(3-methylphenyl)-ethanol (1.7-fold) from 3'-methylacetophenone and (R)-1-(3-methoxyphenyl)-ethanol (1.2-fold) from 3'-methoxyacetophenone, with improved yield. In terms of thermal stability, the ${\underline{M151A}}$/C295A/I86A and ${\underline{V115A}}$/C295A/I86A mutants significantly increased ${\Delta}T_{1/2}$ by $+6.8^{\circ}C$ and $+2.4^{\circ}C$, respectively, with thermal deactivation constant ($k_d$) close to the wild-type enzyme. The ${\underline{M151A}}$/C295A/I86A mutant reacts optimally at $70^{\circ}C$ with almost 4 times more residual activity than the wild type. Considering broad substrate tolerance and thermal stability together, it would be promising to produce (R)-1-(3-methylphenyl)-ethanol from 3'-methylacetophenone by ${\underline{V115A}}$/C295A/I86A, and (R)-1-phenylethanol from acetophenone by ${\underline{M151A}}$/C295A/I86A mutant, in large-scale bioreduction processes.

• 대한수의학회지
• /
• 제28권1호
• /
• pp.199-202
• /
• 1988

The present study was carried out to investigate the serum enzyme activities of racehorses. The enzymes investigated were aspartate and alanine aminotransferase (GOT, GPT), ${\gamma}$-glutamyl transpeptidase(${\gamma}$-GTP), Lactic dehydrogenase(LDH), creatine phosphokinase(CPK), and alkaline phosphatase(ALP). Animals used were 30 healthy racehorses(♀17, ♂13) average weighing 435kg and were from 2 to 8 years of age. LDH activity was progressively decreased with age and next in the order of GOT and CPK activities. GOT and ${\gamma}$-GPT activities were not changed with age but ALP activity tended to be decreased wish age. Activities of GOT, GPT, ${\gamma}$-GTP and ALP were higher in female than in male. There was no difference in CPK activity by sex, and no difference was found out among breeds.

• BMB Reports
• /
• 제28권4호
• /
• pp.363-367
• /
• 1995

E. coli methionyl-tRNA synthetase is one of the class I tRNA synthetases. The Tryptophane residue at the position 461 located in the C-terminal domain of the enzyme is a key amino acid for the interaction with the anticodon of $tRNA^{Met}$. W461 was replaced with other amino acids to determine the chemical requirement for the interaction with the anticodon of $tRNA^{Met}$. Saturation mutagenesis at the position 461 generated a total of 12 substitution mutants of methionyl-tRNA synthetase. All the mutants showed the same in vivo stability as the wild-type enzyme, suggesting that the amino acid substitutions did not cause severe conformational change of the protein The mutants containing tyrosine, phenylalanine, histidine and cysteine substitutions showed in vivo activity while all the other mutants did not. The comparison of the in vitro aminoacylation activities of these mutants showed that aromatic ring structure, Van der Waals volume and hydrogen bond potential of the amino acid residue at the position 461 are the major determinants for the interaction with the anticodon of $tRNA^{Met}$.

• 미생물학회지
• /
• 제20권3호
• /
• pp.105-112
• /
• 1982

The strain of Aspergillus, 6368A, producing acid protease showing high activity was isolated from soil, as a result of wide research about mold group. This strain was identified as a species of Aspergillus tubingensis by the investigation of morphological characteristics. The change of the enzyme production under the various media and culture condition was also studied. The optimum pH and stability of crude acid protease are 2.5, 2.0~4.5 and the optimum temeprature and thermal inactivation waas shown $50^{\circ}C,\;55^{\circ}C$, respectively. From the result of the study on the effects of metal ions, it was found that $MnCl_2,\;CoCl_2,\;CuCl_2,\;SrCl_2,\;and\;NiCl_2$ slightly increased the enzyme activity, on the other hand $ZnCl_2,\;CaCl_2,\;MgCl_2,\;SLS,\;and\;KMnO_4$ decreased it.

• Fisheries and Aquatic Sciences
• /
• 제12권2호
• /
• pp.79-85
• /
• 2009

Gelatin hydrolysates with a high inhibitory activity against angiotensin I-converting enzyme (ACE) were fractionated from Alaska pollock surimi refiner discharge. The ACE-inhibitory activity, expressed as $IC_{50}$ (mg/mL), was highest (0.49 mg/mL) in gelatin hydrolysates formed by sequential 2-hr treatments of Pronase and Flavourzyme. After fractionation through four different membrane filters with molecular weight cut-offs of 3, 5, 10, and 30 kDa, the highest ACE-inhibitory activity (0.21 mg/mL) was observed with the 3-kDa filtrate.

• Journal of Microbiology and Biotechnology
• /
• 제25권2호
• /
• pp.174-177
• /
• 2015

A potent α-glucosidase inhibitor (compound I) was isolated from coffee brews by activity-based fractionation and identified as a β-carboline alkaloid norharman (9H-pyrido[3.4-b]indole) on the basis of mass spectroscopy and nuclear magnetic resonance spectra (¹H NMR, ¹³C NMR, and COSY). The norharman showed potent inhibition against α-glucosidase enzyme in a concentration-dependent manner, with an IC₅₀ value of 0.27 mM for maltase and 0.41 mM for sucrase. A Lineweaver-Burk plot revealed that norharman inhibited α-glucosidase enzyme uncompetitively, with a K_i value of 0.13 mM.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
• /
• 제3권3호
• /
• pp.159-167
• /
• 1999

This study performed experiments for measuring corrosion potential and current density variations in the polarzation curves of polyvinylchloride. The results were examined to identify particular influences affectingthe corrosion potential such as temperature, pH, enzyme, and salt. The lines representing active anodic dissolution were only slightly shifted in the potential direction by temperature, pH, enzyme and salt. The Tafel slope for the anodic dissolution was determined using the polarization effect with varying conditions. The slope of the polarization curves describing the active-to-passive transition region was noticeably shifted in the potential direction. In addition, using the variation in conditions, the best temperature and pH were determined for the corrosion rate, and resistance of corrosion. The second anodic current density peak and maximum passive current density were designated as degraded(IP/I0). The value of IP/I0 was used in measuring the extent of the degradation of the polyvinychloride. The potentiodynamic parameters of the corrosion were obtained using a Tafel plot.

• 자연과학논문집
• /
• 제13권1호
• /
• pp.65-71
• /
• 2003

각종 미강 추출물들의 항고혈압성 엔지오텐신 전환효소 저해 활성을 측정하고 이들의 추출 최적조건을 검토하였다. 각종 미강 추출물들중에서 일품벼 미강의 물 추출물이 77%의 가장 높은 안지오텐신 전환효소 저해활성 보였다. 일품벼 미강중의 안지오텐신 전환효소 저해물질은 물을 1:10(w/v)으로 미강 분말에 첨가한 후 $30^{\circ}C$에서 12시간 추출하였을 때 가장 많이 용출되었다.