• Journal of Microbiology and Biotechnology
• /
• v.17 no.12
• /
• pp.2046-2055
• /
• 2007

Wild-type V. vulnificus cannot grow using lactose as the sole carbon source or take up the sugar. However, prolonged culture of this species in media containing lactose as the sole carbon source leads to the generation of a spontaneous lactose-utilizing (LU) mutant. This mutant showed strong ${\beta}$-galactosidase activity, whereas the wild-type strain showed a barely detectable level of the activity. A mutant with a lesion in a gene homologous to the lacZ of E. coli in the bacterium no longer showed ${\beta}$-galactosidase activity or generated spontaneous LU mutants, suggesting that the lacZ homolog is responsible for the catabolism of lactose, but the expression of the gene and genes for transport of lactose is tightly regulated. Genetic analysis of spontaneous LU mutants showed that all the mutations occur in a lacI homolog, which is located downstream to the lacZ and putative ABC-type lac permease genes. Consistent with this, a genomic library clone containing the lad gene, when present in trans, made the spontaneous LU mutants no longer able to utilize lactose as the sole carbon source. Taken together with the observation that excessive amounts of exogenously supplemented possible catabolic products of lactose have negative effects on the growth and survivability of V. vulnificus, we suggest that V. vulnificus has evolved to carry a repressor that tightly regulates the expression of lacZ to keep the intracellular toxic catabolic intermediates at a sublethal level.

• KSBB Journal
• /
• v.10 no.2
• /
• pp.221-229
• /
• 1995

Effects of carbon sources on zooglan production by Zoogloea ramigera were investigated. The production of zooglan was varied according to the carbon sources used. The largest amount of zooglan was obtained when lactose was used as carbon source and fermentation broth with lactose showed the higher viscosity. The effects of carbon sources were in decreasing order of lactose, glucose, galactose and sucrose. The viscosities of purified zooglan solutions (5g/L) obtained from different carbon sources were measured. When lactose was used, the viscosities of zooglan solutions was quite high and other carbon sources such as glucose and galactose gave little lower viscosities than lactose but sucrose gave very low values. On the other hand, it could be postulated that most of lactose is hydrolyzed by intracellular ${\beta}$-galactosidase.

• KSBB Journal
• /
• v.20 no.1 s.90
• /
• pp.26-33
• /
• 2005

The effects of carbon sources for exopolysaccharide production during batch cultivation of an Enterobacter sp. isolated from the composter were investigated. The highest amount of exopolysaccharide was obtained when lactose was used as carbon source. Lactose in medium was converted into glucose and galactose. Glucose was metabolized fast and was completely consumed, but about $20\%$ of lactose was accumulated as galactose. On the other hand, enzyme activity was about $350\~450$ unit with the increase of lactose concentration. Thus, it was considered that the exopolysaccharide might be produced in the course of that lactose was hydrolyzed into glucose and galactose by $\beta-galactosidase$ with respect to that enzyme activity on lactose hydrolysis was accorded to the exopolysaccharide production. When glucose and galactose were added to lactose medium, respectively, it could be considered that glucose was as a repressor and galactose was as a inducer for $\beta-galactosidase$ synthesis even though the mechanisms were not elucidated. The increase of lactose concentration was almost ineffective to the specific growth rate $(0.133\~0.151\;hr^[-1})$ but showed the difference in the biomass content. The higher carbon source concentration, the more residual sugar remained. It was assumed that the optimum lactose concentration for exopolysaccharide production was $30\~70g/L.$ On the other hand, it was considered that the nitrogen acted as growth limiting nutrients to the cell growth. In the cases of 30 and 70 g/L of the fixed carbon concentrations, the increase of the nitrogen sources concentration caused a remarkable increase within the range of $0.059\~0.225\;hr^{-1}$ and $0.141\~0.237hr^{-1}$ of the specific growth rate, respectively, while there was no significant difference in biomass.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 1979.04a
• /
• pp.113.2-114
• /
• 1979

$\beta-Galactosidase$ is an enzyme which catalizes hydrolysis of lactose, a natural substrate, to glucose and galctose and transferring some monosac-charide units to active acceptors as sugar or alcohol. The occurence of $\beta-Galactosidase$ is known in various microorganisms, animals and higher plants and has been studied by many investigatigators. Especially, a great deal of articles for the enzyme of E. coli have been presented in genetic control mechanism and induction-repression effects of proteins, On the other hand, in the dairly products industry, it is important to hydrolyes lactosd which is the principal sugar of milk and milk products. During the last few years, the interest in enzymatic hydrolysis of milk lactose has teen increased, because of the lactose intolerence in large groups of the population. Microbial $\beta-Galactosidases$ are considered potentially most suitable for processing milk to hydrolyse lactose and, in recent years, the immobilized enzyme from yeast has been examined. Howev, most of the microbial $\beta-Gal$ actosidase are intracellular enzymes, except a few fungal $\beta-Gala-$ ctosidases, and extracellular $\beta-Galactosidase$ which may be favorable to industrial applieation is not so well investigated. On this studies, a mold producing a potent extracellular $\beta-Galactosidase$ was isolated from soil and identified as an imperfect fungus, Beauveria bassians. In this strain, both extracellular and intracellular $\beta-Galactosidases$ were produced simultaneously and a great increase of the extracellular production was acheved by improving the cultural conditions. The extracellular enzyme was purified more than 1, 000 times by procedures including Phosphocellulose and Sephadex G-200 chromatographies. Several characteristics of the enzymewas clarified with this preparation. The enzyme has a main subunit of molecular weight of 80, 000 which makes an active aggregate. And at neutral pH range, it has optimum pH for activity and stability. The Km value was determined to be 0.45$\times$10$^{-3}$ M for $o-Nitrophenyl-\beta-Galactoside.$ In any event, it is interesting to sttudy the $\beta-Galactosidase$ of B. bassiana for the mechanism of secretion and conformational structure of enzyme.

• Food Science and Biotechnology
• /
• v.17 no.3
• /
• pp.514-518
• /
• 2008

After overproduction of a recombinant $\beta$-galactosidase of Bifidobacterium infantis in Pichia pastoris, a synthesis of galacto-oligosaccharides (GOS) from 36% lactose using the enzyme (170.74 U/mg) was investigated. The transgalactosylation ratio reached up to 25.2% with 83.1% conversion of initial lactose and the maximum yield of GOS was 40.6%. The GOS syrup was composed of a 13.43% galacto-oligosaccharides, 5.06% lactose, and 8.76% monosaccharides. The prebiotic effect of GOS on the growth of bifidobacteria and lactobacilli strains was investigated in vitro. The maximum growth rate of Bifidobacterium breve and Lactobacillus acidophillus in GOS syrup (5%, v/v) media were 0.49 and 0.96/hr that are higher than those in 1%(w/v) galactose and 1%(w/v) lactose containing media. However, there was no significant difference between the specific growth rates of L. acidophillus in 1%(w/v) glucose and 5%(v/v) GOS syrup. Our data showed that GOS definitely promoted the growth of B. breve ATCC $15700^T$ and L. acidophilus ATCC 33323.

• Microbiology and Biotechnology Letters
• /
• v.26 no.1
• /
• pp.40-44
• /
• 1998

A Penicillium strain which produces $eta$-galactosidase with high transgalactosylation activity, was isolated from soil and registered as Penicillium sp, KFCC 10888. When $eta$-galactosidase from Penicillium sp. KFCC 10855 reacted with 40% lactose, transgalactosylation ratio reached up to 70% at the 73% conversion of initial lactose. The biosynthesis of the enzyme in Penicillium sp. KFCC 10888 was not induced by lactose. The soybean meal was an effective component of the culture medium. The optimum pH and temperature for transgalactosylation were 4.0 and 55$^{\circ}C$, respectively. The production of galactooligosaccharides was in proportion to the initial lactose concentration. When the enzyme reacted with 40% lactose (pH 4.0) at 55$^{\circ}C$, the concentration of galactooligosaccharides increased up to 40% of total solid concentration.

• Korean Journal of Microbiology
• /
• v.22 no.2
• /
• pp.77-84
• /
• 1984

This examined some conditions for the induction of ${\beta}$-D-galactosidase synthesis in Candida kefyr CBS 834. The optimal pH, temperature, and inoculum size either for growth or${\beta}$-D-galactosidase synthesis were 5.5, $30^{\circ}C$ and above 0.2 at A610nm, respectively. Enzyme activity began to increase at 2h after the addition of inducer, and continued to increase linearly up to $2{\sim}3h$ before reaching stationary phase, and thereafter its activity was decreased. ${\beta}$-D-galactosidase was induced either by lactose or galactose but not either by glucose or ethanol. The greater activity of ${\beta}$-D-galactosidase on galactose than on lactose indicated that the former might be natural inducer for ${\beta}$-D-galactosidase synthesis. The rate of its induction as a function of lactose concentration showed that enzyme activity increased linearly above 4mM, while it was very low below that. Glucose represed the induction of ${\beta}$-D-galactosidase, and the period of adaptation to inducer from other carbon sources was relatively short.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.31 no.2
• /
• pp.202-211
• /
• 2004

Xylitol is a 5-carbons carbohydrate, which can be replaced with sucrose for preventing dental caries. To study the effect of xylitol on the fermentation of lactose in bacteria, the important oral bacteria such as Streptococcus(S.) mutans, S. oralis and S. salivarius were studied. The optical density using spectophotometer and the cell concentration were assessed to evaluate the combined effect of lactose and xylitol against the bacteria. Thin layer chromatography and lactose-PTS activity test were performed to evaluate the effect of xylitol on the fermentation of lactose in S. mutans and by ${\beta}-galactosidase$ with the following results. 1. The optical density of Streptococcus mutans culture was not increased for 8 hours-incubation in the media added with lactose and xylitol, but was increased at 24 hours-incubation. The number of viable cells at 8 hours-incubation was smaller in the media containing lactose and xylitol in comparison with lactose only. 2. The optical densities of Streptococcus oralis culture and Streptococcus salivarius culture were not increased for 8 hours-incubation in the media added with lactose and xylitol but were increased at 24 hours-incubation. 3. When Streptococcus mutars was incubated for 8 hours in the media added with lactose and xylitol, the amount of remained lactose was larger compared with the media added with lactose only But all lactose was fermented in both media after 24 hours-incubation. 4. When Streptococcus mutans was incubated in the media added with lactose and xylitol, the activity of lactose-PTS was higher compared with the media added with lactose only. 5. When ${\beta}-galactosidase$ was incubated in the media added with lactose and xylitol, the amount of remained lactose was larger compared with the media added with lactose only. These results indicated that xylitol inhibited the fermentation of lactose by Streptococcus.

• Microbiology and Biotechnology Letters
• /
• v.18 no.6
• /
• pp.566-570
• /
• 1990

Regulation of $\beta$ -galactosidase formation was studied with Lactobacillus sporogenes. Synthesis of the enzyme was effectively induced by isopropyl- $\beta$-D-thiogalactopyranoside (IPTG) or galactose, and to a much lower level by lactose. When 15 mM glucose was added at the different intervals to the cultures that had been in contact with IPTG, the same levels of inhibition of the enzyme synthesis were observed (approximately one-third the differential rate of a control culture without glucose). This suggests that glucose did not interfere with the entry of the inducer into the cells, but interfere with the formation of $\beta$ -galactosidase through catabolite repression. The glucose inhibitory effect was not overcome by adding CAMP or cGMP to the culture media.

• Microbiology and Biotechnology Letters
• /
• v.9 no.4
• /
• pp.213-218
• /
• 1981

The molecular weight of the purified $\beta$-galactosidase of Penicillium sp. was estimated to be 130000 by both Sephadex G-200 gel filtration and SDS-polyacrylamide del electrophoresis. The SDS-electrophoresis gave two protein bands corresponding to the two molecular weights of 130000 and 70000. These results indicated that the enzyme consisted of two probably identical subunits which had a molecular weight of 70000. The optimum pH of the enzyme activity was 4.7 and maximum activity appeared at 5$0^{\circ}C$. The stable pH range for the enzyme was from 4.5 to 7.0. The purified $\beta$-galactosidase had no metal ion requirement for its activity or stability. The enzyme activity was inhibited by C $u^{++}$(1mM)and galactose (100mM). The hydrolysis of lactose in 5% lactose solution, pasteurized milk and 10% skim milk solution were 69.5%, 88.7% and 72.6% after 4 hr incubation at 5$0^{\circ}C$, when 10 units of $\beta$-glucosidase were used per $m\ell$ of the substrate solutions.s.