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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Journal of Microbiology and Biotechnology
Journal Basic Information
Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 1, Issue 4 - Dec 1991
Volume 1, Issue 3 - Oct 1991
Volume 1, Issue 2 - Aug 1991
Volume 1, Issue 1 - Mar 1991
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Molecular Cloning of
-Galactosidase from Bacillus subtilis HP-4
Kim, Jeong-Ho ; Lee, Jae-Chang ; Huh, Jeong-Won ; Chung, Ki-Chul ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 227~231
A gene coding for a
-galactosidase of Bacillus subtilis HP-4 was cloned in E. coli JM109 by inserting HindIII digested fragment of B. subtilis HP-4 chromosomal DNA into the site of pBR322 and selecting recombinant transformant showing blue color on X-gal plate. The recombinant plasmid, named pBG109, was found to contain the 1.4 Kbp HindIII fragment originated from B. subtilis HP-4 chromosomal DNA by Southern hybridization. The cloned gene was stably maintained and expressed in E. coli JM109 and the pBG109 encoded
-galactosidase had the same enzymatic properties as those of
-galactosidase produced by B. subtilis HP-4.
Purification and In Vitro Translation of Penicillium verruculosum Cellulase mRNA
Kim, Jeong-Ho ; Chung, Ki-Chul ; Kang, Hyun-Sam ; Lee, Young-Kyu ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 232~239
Caboxymethyl cellulase (CMCase) I was purified from the induced culture filtrate of Penicllium verruculosum F-3 by ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Bio-gel P-150 filtration. The purified enzyme was assumed to be a glycoprotein consisting of 8.5% carbohydrate and having a molecular weight of 70.000 in SDS-polycrylamide gel electrophoresis (SDS-PAGE). The purified enzyme-specific anti-CMCase I IgG was obtained by rabbit immunization and protein A-sepharose CL-4B chromatography. The fungal poly(
) RNA was isolated from the total RNA of the mycelium grown under cellulase induction conditions by oligo(dT)-cellulosse chromatography. The translation products in vitro were prepared by translating the isolated poly (
) RNA in rabbit reticulocyte lysate and analyzed by SDS-PAGE and fluorography. Of the translation products, CMCase I was identified by the immunoprecipitation against anti-CMCase I IgG.
A Broad-Host-Range Promoter-Probe Vector, pKU20, and Its Use in Promoter Cloning and Expression of Bacillus thuringiensis Crystal Protein Gene in Pseudomonas putida
SHIN, BYUNG SIK ; BON TAG KOO ; SEUNG HWAN PARK ; HO YONG PARK ; JEONG IL KIM ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 240~245
We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.
Production and Regeneration of Lactobacillus bulgaricus Protoplasts
Jun, Hong-Ki ; Park, Hyun-Jeong ; Baik, Hyung-Suk ; Song, Jae-Chul ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 246~250
Conditions for the production and regeneration in Lactobacillus bulgaricus protoplasts were investigated. Protoplasts of L bulgaricus strains were obtained by treatment with mutanolysin and lysozyme together in a protoplast forming buffer containing 0.02 M N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) (pH 7.0) and 0.5 M sucrose. High protoplast yield was obtained from cells cultured in the de Man, Rogosa and Sharpe(MRS) medium at the middle to late logarithmic growth phase. Regeneration was efficiently accomplished with a complex medium containing 1% sucrose, 20 mM
, 5% gelatin, and 0.5% bovine serum albumin. The frequency of regeneration of protoplasts was 10~20% after 5 days of incubation at
Molecular Cloning and Expression of a Xylanase Gene from Alkalophilic Bacillus sp.
Yu, Ju-Hyun ; Kang, Yun-Sook ; Park, Young-Seo ; Bai, Dong-Hoon ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 251~255
A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.
The Regulation of Alpha-Amylase Synthesis in Bacillus subtilis
Won, Mi-Sun ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 256~260
In B. subtilis,
-amylase synthesis is regulated by amyR located directly on the upstream of amyE. Three different amyR alleles have been reported, amyR1, amyR2 and amyR3. Strains bearing the gra-10 mutation which confers derepression for catabolite repression has GlongrightarrowA transition mutation at ＋5 of amyR1. S1 nuclease mapping demonstrated that transcription initiated at 8 bases downstream from the -10 region of putative E
promoter P1 in amyR1 and gra-10. In amyR2, the major transcription initiatd at the same place and the minor, 10 bases downstream from -10 of P2. The transcript from P2 contributed approximately 15-20% of total amyE mRNA. S1 nuclease protection experiment indicated that amyE mRNA levels corresponded to the rate of synthesis assumed by specific activities of
-amylase in culture supernatants, suggesting that
-amylase synthesis is regulated at the level of transcription.n.
Electron Flow Shift in Clostridium acetobutylicum Fermentation by Lactate
Kwon, Gi-Seok ; Kim, Byung-Hong ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 261~265
Clostridium acetobutylicum produced more butanol in the medium containing corn steep liquor (CSL) than in a complex medium without CSL Addition of CSL to CAB medium increased sugar consumption by the bacterium. Similar results were obtained in the fermentation using CAB medium containing lactate. The ratio for the butanol produced to acetone of the control culture was 1.8, whilst that of the culture containing 44 mM lactate was 5.2. From these results it is hypothesized that lactate functions as an electron flow modulator in the fermentation. This finding has been utilized for the successful butanol fermentation of a non-corn based agricultural byproduct, palm oil waste.
Gene Expression and Secretion of the Anticoagulant Hirudin in Saccharomyces cerevisiae
Sohn, Jung-Hoon ; Lee, Sang-Kwon ; Choi, Eui-Sung ; Rhee, Sang-Ki ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 266~273
Hirudin, a 65-amino acid protein isolated from the salivary gland of the bloodsucking leech, Hirudo medicinalis, is a potent thrombin-specific inhibitor and blocks the thrombin-mediated conversion of fibrinogen to fibrin in clot formation. We have studied the gene expression and secretion of hirudin in yeast. Saccharomyces cerevisiae. A gene coding for hirudin was synthesized based on the amino acid sequence and cloned into a yeast expression vector
factor pre-pro leader sequence and galactose-inducible promoter, GALl0. Recombinant S. cerevisiae was found to secrete biologically active hirudin into the extracellular medium. The secreted recombinant hirudin was recovered from the culture medium and purified with ultrafiltration and reverse phase high performance liquid chromatography. Approximately 1 mg of hirudin per liter was produced under suboptimal culture conditions and brought to about 90% purity in two steps of purification.
Effect of IPTG Induction on Production of
Fusion Protein in Recombinant Escherichia coli
Nam, Soo-Man ; Park, Young-Hoon ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 274~280
Effects of IPTG induction on cell growth and production of
fusion protein (
) were studied in a defined medium using a recombinant Escherichia coli JM109/pCMHB30. IPTG was added (0.2 mM) to induce the cloned-gene expression in the early-, mid-, and late-log growth phases. The most serious decreases in growth rate and plasmid stability were observed for the induction in the early-log growth phase. The expression level of
attained by the induction in the mid-log phase was about 0.51 mg fusion protein/mg total cellular protein, which was 2- and 5-fold improvement over the levels obtained with the inductions in the early- and late-log phases. Formation of acidic byproducts including acetate and pyruvate showed different profiles during the fermentation period for each cases of induction; pyruvate was the major byproduct for the induction in the early-log phase while acetate production became more significant for the cases of inductions in the mid- and late-log phases.
Optimum Conditions for the Formation of Ammonia as a Precursor of Tetramethylpyrazine by Lactococcus lactis ssp. lactis biovar. diacetilactis FC1
Kim, Kyoung-Heon ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 281~284
To investigate the optimum conditions for the production of ammonia as a precursor of tetramethylpyrazine flavor compound from arginine by Lactococcus lactis ssp. lactis biovar. diacetilactis FC1, fermentation factors such as initial pH of culture media, fermentation temperature, concentration of arginine-HC1, and sugars were examined. The optimum conditions were initial pH 5.5 of the culture media, fermentation temperature of
, 6% (w/v) of arginine-HC1, and 1% (w/v) of galactose as a carbon source. Under the optimum fermentation conditions, 40 mmole/l of ammonia was produced after 40 h.
Optimum Conditions for the Formation of Tetramethylpyrazine Flavor Compound by Lactococcus lactis ssp. lactis biovar. diacetilactis FC1
Kim, Kyoung-Heon ; Lee, Hyong-Joo ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 285~287
To produce the tetramethylpyrazine (TMP) flavor compound, Lactococcuss lactis subsp. lactis biovar. diacetilactis (L. diacetilactis) FC1 was cultivated in the TMP medium containing 3% (w/v) of Na-citrate and 6% (w/v) arginine-HC1 as substrates of acetoin and
, respectively, which are the two precursors of the TMP. After 19-day fermentation at
, 0.57 g/l or 4.19 mmole/l of the TMP was produced. This was the first result showing that the TMP could be produced by L. diacetilactis.
Protease Inhibitor Production using Streptomyces sp. SMF13
Kim, In-Seop ; Kim, Hyoung-Tae ; Lee, Hyun-Sook ; Lee, Kye-Joon ;
Journal of Microbiology and Biotechnology, volume 1, issue 4, 1991, Pages 288~292
The aim of the current study is to evaluate the effects of medium compositions on the production of protease inhibitor in Streptomyces sp. SMF13. The production of protease inhibitor was counter-currently linked to extra-cellular protease, which were regulated by the culture conditions. Nitrogen source was the most critical ingredient affecting the production of protease inhibitor and protease. Carbon source was an important factor to determine the culture pH which affected very clearly the formation of protease and protease inhibitor. Inorganic phosphate inhibited the protease inhibitor production which was linked to the cell growth rate, although the optimal conditions for the production of protease inhibitor were not favouring to the cell growth.