• Journal of Microbiology and Biotechnology
• /
• v.24 no.5
• /
• pp.675-682
• /
• 2014

Approximately 50% of people in the world experience abdominal flatulence after the intake of foods containing galactosides such as lactose or soybean oligosaccharides. The galactoside hydrolyzing enzymes of ${\alpha}$- and ${\beta}$-galactosidases have been shown to reduce the levels of galactosides in both the food matrix and the human gastrointestinal tract. This study aimed to optimize the production of ${\alpha}$- and ${\beta}$-galactosidases of Bifidobacterium longum subsp. longum RD47 with a basal medium containing whey and corn steep liquor. The activities of both enzymes were determined after culturing at $37^{\circ}C$ at pH 6.0 for 30 h. The optimal production of ${\alpha}$- and ${\beta}$-galactosidases was obtained with soybean oligosaccharides as a carbon source and proteose peptone no. 3 as a nitrogen source. The optimum pH for both ${\alpha}$- and ${\beta}$-galactosidases was 6.0. The optimum temperatures were $35^{\circ}C$ for ${\alpha}$-galactosidase and $37^{\circ}C$ for ${\beta}$-galactosidase. They showed temperature stability up to $37^{\circ}C$. At a 1 mM concentration of metal ions, $CuSO_4$ inhibited the activities of ${\alpha}$- and ${\beta}$-galactosidases by 35% and 50%, respectively. On the basis of the results obtained in this study, B. longum RD47 may be used for the production of ${\alpha}$- and ${\beta}$-galactosidases, which may reduce the levels of flatulence factors.

• Proceedings of the Korean Society of Life Science Conference
• /
• 2006.09a
• /
• pp.36.2-36.2
• /
• 2006

• Proceedings of the Korean Society of Life Science Conference
• /
• 2006.09a
• /
• pp.36.1-36.1
• /
• 2006

• Proceedings of the Korean Society of Life Science Conference
• /
• 2006.09a
• /
• pp.37.1-37.1
• /
• 2006

• Journal of Microbiology and Biotechnology
• /
• v.8 no.5
• /
• pp.509-516
• /
• 1998

A plasmid vector, pXGPRMATG-lac-Tgx, was developed for overproduction of $\beta$-galactosidase in Escherichia coli using the genetic components of groEx, a heat-shock gene cloned from symbiotic X-bacteria in Amoeba proteus. The vector is composed of intragenic promoters P3 and P4 of groEx, the structural gene of lac operon, transcription tenninator signals of lac and groEx, and ColEl and amp'of pBluescript SKII. The optimized host, E. coli DH5$\alpha$, transfonned with the vector constitutively produced 117,310-171,961 Miller units of $\beta$-galactosidase per mg protein in crude extract. The amount of enzyme in crude extract was 53% of total water-soluble proteins. About 43% of the enzyme could be purified to a specific activity of 322,249 Miller units/mg protein after two-fold purification, using two cycles of precipitation with ammonium sulfate and one step of gel filtration. Thus, the expression system developed in this study presents a low-cost and simple method for purifying overproduced $\beta$-galactosidase in E. coli.

• Journal of Plant Biotechnology
• /
• v.43 no.2
• /
• pp.164-173
• /
• 2016

To investigate the genetic basis of phenotypic differences, sequence variations were analyzed in 8 inbred watermelon lines by re-sequencing. The number of sequence variations differed depending on the chromosome. Only 12.9% of SNPs were found within genes, whereas the rest were detected in promoter or intergenic regions. SNP density analysis showed that there was a highly variable region at the end of chromosome 6, which is similar to previously published findings. However, this region with high SNP density did not show much variation between the lines. In contrast, highly conserved regions with a size of 6.5-10 Mb were found in chromosomes 10 and 11. Pathway analysis suggested that the DIMBOA (a natural antibiotic)-glucoside degradation pathway was significantly different between the lines, indicating that the eight lines may have different levels of pathogen resistance. Among the carbohydrate-related genes, the alpha-galactosidase gene was the most variable among the lines. Information from this study will be helpful in understanding the watermelon breeding process at the molecular level.

• Journal of Genetic Medicine
• /
• v.15 no.1
• /
• pp.28-33
• /
• 2018

Fabry disease (FD) is a rare X-linked lysosomal storage disorder caused by the deficiency of ${\alpha}$-galactosidase A. Patients with classical FD present acroparesthesia, hypohidrosis, cornea verticillata, disseminated angiokeratoma, and microalbuminuria in childhood, and develop life-threatening renal, cardiac, and cerebrovascular complications typically after the fourth decade of life. To date, more than 700 mutations responsible for FD have been identified in the human GLA gene. Herein, we report a novel GLA mutation, c.1117_1141del25 (p.Gly373Profs*10), identified in an 11-year-old Korean boy with FD presenting early cardiac and neurologic manifestation and in other affected family members. The boy had acroparesthesia, hypohidrosis, cornea verticillata, and left ventricular hypertrophy. His mother and sister also had acroparesthesia. Two males on the mother's side had similar pain and died of unknown causes. The plasma ${\alpha}$-galactosidase A activity (4.1 nmol/hr/mg protein) of the patient was markedly lower than the mean value of the controls. The plasma level of globotriaosylsphingosine was elevated in the patient and all the carriers. We concluded the novel GLA mutation c.1117_1141del25 is a pathogenic mutation for FD, probably related to the early cardiac manifestation of FD.

• Journal of the Korean Society of Radiology
• /
• v.81 no.2
• /
• pp.302-309
• /
• 2020

Fabry disease is a rare X-linked metabolic disorder that is characterized by the accumulation of glycosphingolipids in various organs, resulting from the deficiency of alpha-galactosidase A. Cardiac involvement is relatively common; myocardial inflammation, left ventricular hypertrophy, and myocardial fibrosis secondary to abnormal lipid deposition in myocytes are often observed. Hence, the diagnosis of cardiac involvement is crucial for evaluating patient prognosis. Cardiac MRI is the standard technique for measuring the function, volume, and mass of the ventricles. It is also useful for myocardial tissue characterizations. The evaluation of native myocardial T1 values can facilitate early diagnosis of cardiac involvement, while measurements of left ventricular myocardial mass can be used to monitor treatment outcomes, in patients with Fabry disease. Consequently, cardiac MRI can provide useful information for diagnosing, monitoring, and treating patients with Fabry disease.

• Microbiology and Biotechnology Letters
• /
• v.25 no.5
• /
• pp.448-453
• /
• 1997

Enzyme activities, production of killer toxin and some functionality of forty seven yeasts isolated from traditional Meju were investigated in culture broth and cell free extracts. Activities of $\alpha$-galactosidase, invertase and inulinase were detected in cell free extracts of 38 strains, 43 strains and 45 strains, respectively and acidic and neutral protease activities also were detected in culture broth of all the strains, $\beta$-Galactosidase activity was detected in cell free extracts of OE-20 and S-14 strains. Killer toxins were produced by OE-12, S-8 (Candida spp.), OE-19 (Zygosaccharomyces spp.) and S-3 (Saccharomyces spp.). Culture broth of OE-23 and S-9 showed 61.3% and 59.2% of antioxidant activity to $\alpha$, $\alpha$-diphenyl-$\beta$-picrylhydrazyl(DPPH), but nitrite-scavenging ability as well as inhibition of tyrosinase and polyphenol oxidase were not appeared in all the strains.

• Journal of Microbiology
• /
• v.35 no.1
• /
• pp.61-65
• /
• 1997

A novel strategy was developed for isolating suppressors from sporulation-deficient mutants. The mutation in the BCY1 gene, which codes for the regulatory subunit of cAMP-dependent protein kinase, when homozygous, results in diploids being meiosis and sporulation deficient. Two plasmids, YCp-MAT.alpha. and YEp-SPOT7-lacZ, were introduced into MAT.alpha. BCY1$\^$+/ or MAT.alpha. bcy1 haploid cells. The transformant of the BCY1$\^$+/ haploid cell produced .betha.-galactosidase under nutrient starvation, but the bcy1 transformant did not. Using this system, the mutagenesis experiment performed on the bcy1 transformant strain resulted in a number of sporulation mutants that produced .betha.-galactosidase under nutrient starvation. One complementation group, sob1, was identified from the isoalted suppressor mutants and characterized as a single recessive mutation by tetrad analysis. Genetic analysis revealed that the sob1 mutation suppressed the sporulation deficiency, the failure to arrest at the G1 phase of the cell cecle, and the sensitivity to heat or nitrogen starvation caused by the bcy1 mutation. However, the sob1 mutation did not suppress the sporulation deficiency of ime1 and of ime2 diploids. These results suggest that the sob1 mutation affects a gene which functions as a downstream regulator in both meiosis and cell cycle regulation.