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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 4, Issue 4 - Dec 1994
Volume 4, Issue 3 - Sep 1994
Volume 4, Issue 2 - Jun 1994
Volume 4, Issue 1 - Mar 1994
Selecting the target year
Detection and Determination of Lipase Activity
Lee, Seoung-Yong ; Rhee, Joon-Shick ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 85~94
Lipase (triacylglycerol hydrolase, EC 22.214.171.124) is able to catalyze the hydrolysis of ester bonds of triacylglycerols at the interface between aqueous phase and organic phase containing substrate. With the rapid development of lipid biotechnology, lipase-catalyzed hydrolysis of lipids has a great concern from the industrial point of view. Owing to the reversible nature of the lipase, the reactions are also applied for glyceride synthesis, interesterification and resolution of racemic mixtures into optically active alcohols or acids. For all applications of the lipases, a reliable method for the determination of enzyme activity is required. Precise quantitative determination of its activity is essential as the basis of research and development of the bioprocess involving the enzyme. This article reviews the existing literature on the detection and determination of lipase activity from microbial, mammalian and plant sources.
Effect of Glycine on L-Ornithine Production by a Citrulline Auxotroph of Brevibacterium ketoglutamicum and Stoichiometric Analysis
Nam, Soo-Wan ; Choi, Dae-Keon ; Ryu, Wuk-Sang ; Jang, Hyung-Wook ; Chung, Bong-Hyun ; Park, Young-Hoon ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 95~101
The effects of glycine on cell growth and L-omithine production were investigated in shake-flask and jar fermentor cultures of a citrulline auxotrophic mutant, Brevibacterium ketoglutamicum BK 1046. In the shake-flask culture, the optimal concentration of glycine for L-ornithine production was found to be 20 g/l. In the jar fermentor culture with the glycine at an initial concentration of 20 g/l, L-ornithine production increased by 28%, compared to that of the culture with no glycine added. 37 g/l of L-ornithine was produced when additional feeding of glycine (5 g/l) was made. This was a significant improvement in L-ornithine production compared to that (ca. 24 g/l) of the corresponding batch culture conducted without glycine. According to the stoichiometric analysis with the batch fermentation results, the experimental and theoretical L-ornithine yields based on the glucose consumption were 0.24 and 0.59, respectively. This indicates that the performance of L-ornithine fermentation can further be improved by the supplementation of glycine and the development of a mutant strain possessing a higher growth yield.
Cloning of the Endoglucanase Gene from Actinomyces sp. 40 in Escherichia coli and Some Properties of the Gene Products
Min, Hae-Ki ; Choi, Yun-Jaie ; Cho, Kwang-Keun ; Ha, Jong-Kyu ; Woo, Jung-Hee ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 102~107
-1,4-endoglucanase gene from Actinomyces sp. 40 was cloned into Escherichia coli DH5
with pUC19. Chromosomal DNA from Actinomyces sp. 40 was cleaved with the restriction enzyme Sau3AI and ligated into pUC19 for the transformation of Escherichia coli DH5
. Positive clones of
-1,4-endoglucanase gene were detected as the clear zones on a medium supplemented with carboxymethylcellulose (CMC). This transformant possessed a single plasmid, designated pDS1, which contained the vector DNA and a 3.5 kilobase (kb) Sau3AI insertion fragment encoding endoglucanase. The size of the cloned fragment was reduced to 2.0 kb. The endoglucanase activity produced by the E. coli DH5
(pDS6) was higher than that of Actinomyces sp. 40 strain. The optimum pH and temperature of the cloned enzyme were pH 4.0
5.0 and 55
, respectively. The cloned enzyme was stable at 55
or below and in buffer ranging from pH 4.0 to 7.0. The enzyme degraded CMC but did not degrade xylan, cellobiose, and methyl-umbelliferylcellobiopyranoside (MUC).
Genetic Organization of the Recombinant Bacillus pasteurii Urease Genes Expressed in Escherichia coli
Kim, Sang-Dal ; Hausinger, Robert P. ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 108~112
The genetic organization of the urease gene cluster from an alkalophilic Bacillus pasteurii was determined by subcloning and Tn5 transposon mutagenesis of a 10.7 kilobasepair cloned fragment. A region of DNA between 5.0 and 6.0 kb in length is necessary for urease activity. In vitro transcription-translation analysis of transposon insertion mutants of the cloned urease genes demonstrated that the major (
67,000) and minor (
20,000) structural peptides of urease are encoded at one end of the urease gene cluster and at least 3 additional polypeptides are encoded by adjacent DNA sequences.
Purification and Characterization of a Thermostable Protease from Pseudomonas aeruginosa NS-83
Kim, Hyung-Kwoun ; Kim, Kee-Hyun ; Lee, Jung-Kee ; Bae, Kyung-Sook ; Sung, Chang ; Oh, Tae-Kwang ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 113~118
A bacterial strain NS-83 isolated from soil was able to produce an extracellular thermostable protease. The strain was identified as Pseudomonas aeruginosa based on its morphological and physiological characteristics. A thermostable protease from this strain has been purified to homogeneity as judged by SDS-PAGE and isoelectric focusing. The purification procedures included hydrophobic interaction, ion exchange, and gel filtration chromatography. The
and the pl of the enzyme were 32,000 and 5.9, respectively. The optimal pH at 55
and the optimal temperature at pH 7.0 were 8.0 and 60
, respectively. The D-values of the enzyme at 60, 65, and 70
were 22, 2.1, and 0.75 hrs, respectively. The enzyme activity was significantly inhibited in the presence of 1 mM o-phenanthroline or EDTA, suggesting that the enzyme is metalloprotease. The
for Hammarsten casein were found to be 3.2 mg/ml and 0.918 unit/ml, respectively. These enzymatic properties were similar to those of elastase produced from P. aeruginosa IFO 3455, but the enzyme was clearly different from the reported elastase, in respect to
effects on enzyme-thermostability. This property, together with amino acid composition analysis, confirmed that the enzyme differs from the known P. aeruginosa elastase.
Enzymatic Characteristics of steroid
-dehydrogenase from Arthrobacter simplex
Lee, Mi-Kyung ; Bae, Moo ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 119~125
-dehydrogenase purified from hydrocortisone-induced cells of Arthrobacter simplex converted various 3-ketosteroids into their corresponding
-dehydrogenated products. The transformation efficiencies depend upon the chemical structure of the steroids, especially length of the side chain at 17 position and hydroxyl groups at 11 and 17 positions. The Km values for androstenedione, the most favorable substrate examined, and hydrocortisone were 74
, respectively. The optimum temperature and pH of the enzyme reaction were 35
and pH 9, respectively, and the enzyme was relatively stable at the range from 20 to 35
and from pH 5 to 10 after one hour of incubation. The enzyme activity was markedly inhibited in the presence of
ions, and somewhat inhibited by
-Dipyridyl that inhibits 9
-hydroxylase and accumulates 1,4-androstadiene-3,17-dione from sterols revealed no inhibitory effect on this enzyme. EGTA showed inhibitory effect.
-Estradiol competitively inhibited the enzyme activity. Chemical modifications of the enzyme were attempted with several reagents. p-Hydroxymer-curibenzoate showed inhibition of the enzyme activity and protection of the substrate. This suggests that cysteine residue may be involved in the active site of the enzyme.
Isolation of Pseudomonas putida BM01 Accumulating High Amount of
Song, Jae-Jun ; Yoon, Sung-Chul ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 126~133
A Pseudomonas putida strain able to accumulate high amount of polyesters of medium-chain-length 3-hydroxyalkanoic acids (
) was isolated from soil in a landfill site using an enrichment technique. Culture condition of the isolated strain for polyester production in a one-step culture was optimized in a mineral-salts medium against pH and concentrations of ammonium sulfate, carbon source(e.g., octanoate), and phosphate. The optimal values for maximal cell growth and PHA accumulation were: pH; 7
; 8 mM, octanoate; 40 mM. The optimum temperature was in the range of
, which was rather broader than in other bacteria. Cell growth was strongly inhibited by the phosphate limitation to less than 1 mM. An increase of phosphate concentration above 1 mM showed little effect on cell growth and polyester accumulation. When the strain was grown on octanoate under this optimized condition it produced 3.4 g dry biomass per liter and yielded 1.7 g PHA per liter amounting to 53 wt% of dry cells. The monomer units composing the polyester synthesized from octanoate were 3-hydroxyoctanoate (3HO), 3-hydroxycaproate (3HC), and 3-hydroxybutyrate (3HB) (85:13:2, mole ratio). Other low linear
monocarboxylic acids were also tested for polyester production.
The Production and Enzymatic Properties of Extracellular Chitinase from Pseudomonas stutzeri YPL-1, as a Biocontrol Agent
Lim, Ho-Seong ; Kim, Sang-Dal ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 134~140
An antagonistic bacterium Pseudomonas stutzeri YPL-1 liberated extracellular chitinase and
-1,3-glucanase which are key enzymes in the decomposition of fungal hyphal walls. The lytic enzymes caused abnormal swelling and retreating at the hyphal tips of plant pathogenic fungus Fusarium solani in a dual culture. Scanning electron microscopy revealed the hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. The production of chitinase and properties of a crude preparation of the enzyme from P. stutzeri YPL-1 were investigated. Peak of the chitinase activity was detected after 4 hr of cultivation. The enzyme had optimum temperature and pH of 50
and pH 5.3, respectively. The enzyme was stable in the pH range of 3.5 to 6.0 up to 50
. The enzyme was significantly inhibited by metal compounds such as
, but was stimulated by
. P. stutzeri YPL-1 produced high levels of the enzyme after 84 hr of incubation. Among the tested carbon sources, chitin was the most effective for the enzyme production, at the concentration level of 3%. As a source of nitrogen, peptone was the best for the enzyme production, at the concentration level of 4%. The maximum amount of enzyme was produced by cultivating the bacterium at a medium of initial pH 6.8.
Use of Nisin as an Aid in Reduction of Thermal Process of Bottled Sikhae (Rice Beverage)
Yoo, Jin-Young ; Kwon, Dong-Jin ; Park, Jong-Hyun ; Koo, Young-Jo ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 141~145
Conventional commercial thermal process for preparing Sikhae (Rice beverage) in a hermetically sealed container was evaluated to solve the nutritional deterioration and organoleptic inferiority problem caused by severe heat treatment. A milder thermal process with an aid of Nisin, a GRAS-grade, selectively germicidal compound, was introduced to destroy the putrefactive microorganisms. In this experiment, hot-filling method with Nisin, and thermal processing (at 110
for 15 minutes with Nisin, at 121
for 25 minutes without Nisin) were compared. The quality of Sikhae could be enhanced and over 90% of the thermal process could be conserved by this process in terms of sterilizing value without quality deterioration when processing the bottled Sikhae at 110
for 15 minutes
Optimization of Fed-Batch Fermentation for Production of Poly-
-Hydroxybutyrate in Alcaligenes eutrophus
Lee, In-Young ; Choi, Eun-Soo ; Kim, Guk-Jin ; Nam, Soo-Wan ; Shin, Yong-Cheol ; Chang, Ho-Nam ; Park, Young-Hoon ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 146~150
Production of poly-
-hydroxybutyrate (PHB) in fed-batch fermentation was studied. Utilization of carbon for PHB biosynthesis was investigated by using feeding solutions with different ratios of carbon to nitrogen (C/N). It was observed that at a high C/N ratio carbon source was more preferably utilized for PHB accumulation while its consumption for cellular metabolism appeared to be more favored at a low C/N value. A high cell concentration (184 g/l) was achieved when ammonium hydroxide solution was fed to control the pH, which was also utilized as the sole nitrogen source. For the mass production of PHB, two-stage fed-batch operations were carried out where PHB accumulation was observed to be stimulated by switching the ammonium feeding mode to the nitrogen limiting condition. A large amount of PHB (108 g/l) was obtained with cellular content of 80% within 50 hrs of operation.
Selection of Human-Originated Lactobacillus acidophilus For Production of Probiotics
Kim, Wang-June ; Hong, Seok-San ; Cha, Seong-Kwan ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 151~154
Lactobacillus acidophilus KFRI 233, a strain isolated from human, was selected as a candidate for probiotics due to its excellent growth in MRS broth where no special anaerobic condition is required. Both simultaneous and deferred agar diffusion assays exhibited Lb. acidophilus KFRI 233 to possess an antagonistic effect against Clostridium perfringens. Its antagonistic effect was pH dependent Associative culture of KFRI 233 and Cl. perfringens in broth resulted in maximum 94.04% inhibition of Cl. perfringens.
-Galactosidase activity of KFRI 233 was higher than other tested strains that are sold as commercial probiotics. Survival of KFRI 233 in pasteurized skim milk (4
) and Sherbet mix (-15
) after 7 days of storage were 71.9 and 105.5%, respectively.
Expression of lac and gal operons in Zymomonas mobilis
Cho, Dong-Wuk ; Rogers, Peter L. ; Delaney, Stephen F. ;
Journal of Microbiology and Biotechnology, volume 4, issue 2, 1994, Pages 155~159
Two Zymomonas mobilis strains (ZM63 and ZM6307), containing both lactose and galactose operons, were constructed.
-Galactosidase and galactokinase assays indicated that both operons were expressed in both strains. The transport systems available for lactose uptake by Zymomonas mobilis were investigated using
-labelled lactose. After the outer membrane, which was considered to be a possible barrier to lactose uptake, was disrupted by treatment with EDTA and
ions, some increase in lactose uptake was observed in ZM6306 (
) and ZM6307 (
), but not in the parent, ZM6. This suggested that the outer membrane of Zymomonas mobilis acts as a barrier to lactose uptake to some degree, and also that the lactose permease is operational in Zymomonas mobilis.