• Korean Journal of Food Science and Technology
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• v.28 no.3
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• pp.451-457
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• 1996

The effects of pH and L-cysteine HCI on the growth and stability of Biofidobacterium breve were studied. Significantly higher population was obtained by culturing at pH $6.0{\sim}6.5$ than at any other pH. The cultures that had been grown at pH $5.5{\sim}6.0$ were more stable during storage than those grown at other pH. The number of B. breve that had been grown at pH 5.5 and 6.0 remained as $2.4{\times}10^6ml/\;and\;1.4{\times}10^6ml,$ respectively, after 25 days of storage at $4^{\circ}C$. The ${\beta}$-galactosidase activity of B. breve grown at pH 5.5 and 6.0 was reduced only to $78{\sim}85%$ of the control after the same storage condition, whereas the culture grown at pH 7.0 exhibited a signficant decline in population and ${\beta}$-galactosidase activity during $4^{\circ}C$. The growth of B. breve was promoted by 0.05% L-cysteine HCI, and cells grown in MRS with $0.05{\sim}0.10%$ L-cysteine HCI were more resistant to hydrogen peroxide. With respect to the effect to the effect of osmoprotectants on the survival of B. breve subjected to freeze-drying, addition of 2 mM betaine of 2 mM trehalose increased the growth rate of cells grown under osmotic stress and also made the organism more osmotolerant. Furthermore, the betaine or trehalose increased the survivability of the cells after freeze-drying.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1392-1400
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• 2017

Galactooligosaccharides (GOSs) are known to be selectively utilized by Bifidobacterium, which can bring about healthy changes of the composition of intestinal microflora. In this study, ${\beta}-GOS$ were synthesized using bifidobacterial ${\beta}-galactosidase$ (G1) purified from recombinant E. coli with a high GOS yield and with high productivity and enhanced bifidogenic activity. The purified recombinant G1 showed maximum production of ${\beta}-GOSs$ at pH 8.5 and $45^{\circ}C$. A matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the major peaks of the produced ${\beta}-GOSs$ showed MW of 527 and 689, indicating the synthesis of ${\beta}-GOSs$ at degrees of polymerization (DP) of 3 and DP4, respectively. The trisaccharides were identified as ${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-glucopyranose, and the tetrasaccharides were identified as ${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-glucopyranose. The maximal production yield of GOSs was as high as 25.3% (w/v) using purified recombinant ${\beta}-galactosidase$ and 36% (w/v) of lactose as a substrate at pH 8.5 and $45^{\circ}C$. After 140 min of the reaction under this condition, 268.3 g/l of GOSs was obtained. With regard to the prebiotic effect, all of the tested Bifidobacterium except for B. breve grew well in BHI medium containing ${\beta}-GOS$ as a sole carbon source, whereas lactobacilli and Streptococcus thermophilus scarcely grew in the same medium. Only Bacteroides fragilis, Clostridium ramosum, and Enterobacter cloacae among the 17 pathogens tested grew in BHI medium containing ${\beta}-GOS$ as a sole carbon source; the remaining pathogens did not grow in the same medium. Consequently, the ${\beta}-GOS$ are expected to contribute to the beneficial change of intestinal microbial flora.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.437-442
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• 1996

Two strains of Lactobacillus(L.) casfi and one strain of L. Pentosus, which were isolated from pickles, were used to investigate in studing their characteristics of ${\beta}-galactosidase$. The preferable carbon sources and pH of the MRS media for enzyme production from L. casei No.10 was found to be 1.0% lactose and pH 7.5, from L. Pentosus No.63 was 1.0% galactose and pH 7.5, and from L. casei No.36 was 1.0% lactose and pH 6.5, respectively. The maximum enzyme production from each strain was found after 48 hours culture at $30^{\circ}C$ in a medium with preferable carbon source. The optimum reaction temperature with substrate for ${\beta}-galactosidase$ activity was found at $60^{\circ}C$ for all three strains . The stability of enzyme from L. casei No.36 was found to be at $45^{\circ}C$, from L. Pentosus No.63 was found at $55^{\circ}C$. This stability from L. casei No.36 was found at $40^{\circ}C$, but it was reduced to 60% at $55^{\circ}C$. These stabilities of enzymes remained about 90% at $40^{\circ}C$ for all three strains. The optimal pH for enzyme activities was found to be pH 6.5 for all three strains. Enzyme activity remained over 90% for L. casei No.10 at $pH\;5.0{\sim}6.0$, for L. casei No.36 at $pH\;5.0{\sim}8.0$, and for L. pentosus No.63 at $pH\;6.0{\sim}7.0$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.29-34
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• 1998

This study was investigated to know changes of the cell wall components, cell wall degrading enzyme activities and contents of soluble protein of strawberry during ripening and softening. The contents of water soluble substances were slightly increased during ripening, but the contents of alcohol-insoluble substances were not changed. The contents of pectin were not changed at green mature and turning stage, while decreased after mature stage. The contents of alkali-soluble hemicellulose and cellulose were increased during ripening and softening. The contents of water-soluble and saltsoluble protein were not changed, but the content of cell wall protein was slightly decreased during ripening. The content of total protein was increased at turning stage, it is not changed after turning stage. $\beta$-Galactosidase activity was increased during ripening, and pectinmethylesterase activity was decreased at turning. Phenylalanine ammonia-lyase activity was changed up to mature stage, but decreased at overripening stage. Polygalacturonase and cellulase activities were not detected at all of ripening stages.

• Journal of Sericultural and Entomological Science
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• v.39 no.1
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• pp.36-43
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• 1997

In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.287-294
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• 2008

Three amino acid residues (His119, Glu164, and Glu338) in the active site of Thermus caldophilus GK24 ${\beta}$-glycosidase (Tca ${\beta}$-glycosidase), a family 1 glycosyl hydrolase, were mutated by site-directed mutagenesis. To verify the key catalytic residues, Glu164 and Glu338 were changed to Gly and Gln, respectively. The E164G mutation resulted in drastic reductions of both ${\beta}$-galactosidase and ${\beta}$-glucosidase activities, and the E338Q mutation caused complete loss of activity, confirming that the two residues are essential for the reaction process of glycosidic linkage hydrolysis. To investigate the role of His119 in substrate binding and enzyme activity, the residue was substituted with Gly. The H119G mutant showed 53-fold reduced activity on 5mM p-nitrophenyl ${\beta}$-D-galactopyranoside, when compared with the wild type; however, both the wild-type and mutant enzymes showed similar activity on 5mM p-nitrophenyl ${\beta}$-D-glucopyranoside at $75^{\circ}C$. Kinetic analysis with p-nitrophenyl ${\beta}$-D-galactopyranoside revealed that the $k_{cat}$ value of the H119G mutant was 76.3-fold lower than that of the wild type, but the $K_m$ of the mutant was 15.3-fold higher than that of the wild type owing to the much lower affinity of the mutant. Thus, the catalytic efficiency $(k_{cat}/K_m)$ of the mutant decreased to 0.08% to that of the wild type. The $k_{cat}$ value of the H119G mutant for p-nitrophenyl ${\beta}$-D-glucopyranoside was 5.l-fold higher than that of the wild type, but the catalytic efficiency of the mutant was 2.5% of that of the wild type. The H119G mutation gave rise to changes in optima pH (from 5.5-6.5 to 5.5) and temperature (from $90^{\circ}C\;to\;80-85^{\circ}C$). This difference of temperature optima originated in the decrease of H119G's thermostability. These results indicate that His119 is a crucial residue in ${\beta}$-galactosidase and ${\beta}$-glucosidase activities and also influences the enzyme's substrate binding affinity and thermostability.

• Korean Journal of Food Science and Technology
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• v.22 no.2
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• pp.129-133
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• 1990

The strain of Nuruk yeat No. 15 (NY-15) which ferments lactose in milk was Isolated from Nuruk and identified as Saccharomyces marxianus according to the API 20C profile index. The lactose hydrolysing ability of NY-15 was similar to that Saccharomyces fragilis ATCC 8583 which has ${\beta}-galactosidase$ activity. Its optimum growth temperature, pH and time for the production of maximum enzyme activity showed $28^{\circ}C$, 4.5 and 28hr, respectively. Galactose as well as sucrose as carbon sources, and urea as nitrogen source Increased the production of enzyme. In order to test the production of alcohol, NY-15 was inoculated in whey medium and whey medium added with sugar. In the former, NY-15 produced 2% alcohol and in the latter, it showed 12% alcohol production. The optimal medium pH for lactose hydrolysis of NY-15 is 4.5, whereas that of Saccharomyces fragilis ATCC 8583 is 3.5

• Applied Biological Chemistry
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• v.38 no.1
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• pp.7-12
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• 1995

The purpose of this study was the development of promoter for the lacZ' gene. Two heterologous promoter I and II of lacZ' gene were isolated from chromosomal DNA Bam HI fragment of yeast. The size of the promoter I was estimated to be 2.5 kb and ${\beta}-galactosidase$ activity was 124.6 U/mg protein, and the size of the promoter II was 4.0 kb and its ${\beta}-galactosidase$ activity was 168.8 U/mg protein, respectively. The stability of the recombinant YEp plasmid in the transformant was from 52.7 to 67.4% at minimal medium. YIp plasmid was constructed from YEp plasmid, and expressed both in E. coli and yeast. The promoter I aid II iso-lated from yeast chromosomal DNA can be used for promoter of plasmid YEp and YIp.

• Journal of Microbiology and Biotechnology
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• v.4 no.2
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• pp.151-154
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• 1994

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. $\beta$-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$^{\circ}C$) and Sherbet mix (-15$^{\circ}C$) after 7 days of storage were 71.9 and 105.5%, respectively.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.550-556
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• 1995

The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.