• Journal of Microbiology
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• v.42 no.4
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• pp.319-327
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• 2004

An additional amylase, besides the typical $\alpha-amylase,$ was detected for the first time in the cytoplasm of B. subtilis SUH4-2, an isolate from Korean soil. The corresponding gene (bbmA) encoded a malto­genic amylase (MAase) and its sequence was almost identical to the yvdF gene of B. subtilis 168, whose function was unknown. Southern blot analysis using bbmA as the probe indicated that this gene was ubiquitous among various B. subtilis strains. In an effort to understand the physiological function of the bbmA gene in B. subtilis, the expression pattern of the gene was monitored by measuring the $\beta-galactosidase$ activity produced from the bbmA promoter fused to the amino terminus of the lacZ struc­tural gene, which was then integrated into the amyE locus on the B. subtilis 168 chromosome. The pro­moter was induced during the mid-log phase and fully expressed at the early stationary phase in defined media containing $\beta--cyclodextrin\;(\beta-CD),$ maltose, or starch. On the other hand, it was kept repressed in the presence of glucose, fructose, sucrose, or glycerol, suggesting that catabolite repression might be involved in the expression of the gene. Production of the $\beta-CD$ hydrolyzing activity was impaired by the spo0A mutation in B. subtilis 168, indicating the involvement of an additional regu­latory system exerting control on the promoter. Inactivation of yvdF resulted in a significant decrease of the $\beta-CD$ hydrolyzing activity, if not all. This result implied the presence of an additional enzyme(s) that is capable of hydrolyzing $\beta-CD$ in B. subtilis 168. Based on the results, MAase encoded by bbmA is likely to be involved in maltose and $\beta-CD$ utilization when other sugars, which are readily usable as an energy source, are not available during the stationary phase.

• Korean Journal of Poultry Science
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• v.29 no.1
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• pp.19-23
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• 2002

This studs was conducted to investigate the effects of dietary carbohydrase (multi-enzyme: $\alpha$-galactosidase and mannanase) on egg quality and nutrient digestibility in laying hens. One hundred forty four, 47-wk-old, ISA Brown commercial layers were used in a 28-d feeding trial after a 7-d adjustment period. Dietary treatments were 1) CON(basal diet), 2) ME 0.1 (basal diet +0.1% multi -enzyme), 3) ME 0.2 (basal diet + 0.2% multi-enzyme). Fer overall Period, hen-day egg Production, egg weight, egg shell breaking strength and egg shell thickness were not influenced by the multi-enzyme. As the adding levels of multi-enzyme increased in the diet, egg Yolk color and egg Yolk index tended to increase with significant differences. Digestibility of DM was not affected by multi-enzyme. However, digestibility of N increased significantly as the concentration of multi-enzyme in the diet was increased. In conclusion, supplemental carbohydrase in laying hen diets nay have some roles in improving the egg Yolk color and N digestibility.

• Korean Journal of Agricultural Science
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• v.41 no.4
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• pp.265-274
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• 2014

Physiological characteristics of two Korean golden kiwifruit cultivars, 'Halla Gold' and 'Haehyang', were compared to determine the storage potential of fruit. The soluble solid levels of the fruit were 8.9 and 6.9 oBrix in 'Halla Gold' and 'Haehyang' at harvest, respectively but increased up to 15.4 in 'Halla Gold' and 17.5 oBrix in 'Haehyang' after 2 months of storage. Major sugars were fructose and glucose, and sucrose content was relatively low regardless of cultivar. The edible quality of 'Haehyang' was better than 'Halla Gold' because of higher amount of sugars. Firmness of the fruits gradually decreased as the increase of storage period in 'Halla Gold' in both flesh and core tissue. Th firmness loss of 'Haehyang' fruit was faster in the first 2 months and then became slow. After 75 days of storage, the firmness of 'Haehyang' fruit was only 5.2% at harvest. Core tissue was soften enough to eat at ripe stage. Wall modifying enzyme activities including xylanase, ${\alpha}$-L-arabinofuranosi-dase and ${\beta}$-galactosidase were consistently higher in 'Haehyang' and the activity of pectate lyase was more increased than 'Halla Gold' after 2 months of storage. Respiration rate of 'Haehyang' was higher than 'Halla Gold' and further increased after 2 months of storage. Weight loss was much higher in 'Haehyang' which showed higher rate of the firmness loss. The storage potential of golden kiwifruit was estimated to be about 2 months for 'Haehyang' and 3 months for 'Halla Gold' when determined on the basis of the fruit firmness.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.749-755
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• 2005

The ccpA gene encoding catabolite control protein A (CcpA) of Leuconostoc mesenteroides SYl, a strain isolated from kimchi, was cloned, sequenced, analyzed for transcript, and overexpressed in Escherichia coli. The ccpA ORF (open reading frame) is 1,011 bp in size, which can encode a protein of 336 amino acid residues with a molecular mass of 36,739 Da. The transcription start site was mapped at a position 49 nucleotides upstream of the start codon, and promoter sequences were also identified. The putative cre site overlapped with the -35 promoter sequence. The deduced amino acid sequence of the CcpA contained the helix-turn-helix motif found in many DNA-binding regulatory proteins. CcpA from 1. mesenteroides SY1 had $54.6\%$ identity with CcpA from Lactobacillus casei. The Northern blot experiment showed that ccpA was transcribed as a single 1.1 kb transcript, and transcription was repressed when grown on media containing glucose. CcpA was overproduced in E. coli BL21(DE3) cells using the pET expression vector, and purified to an apparent homogeneity. Gel Mobility Shift Assay with purified CcpA and a DNA fragment containing the ere sequence of the $\alpha$-galactosidase gene (aga) from L. mesenteroides SY1 revealed that CcpA bound specifically to the cre site of aga.

• Journal of Nutrition and Health
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• v.35 no.9
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• pp.912-918
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• 2002

In nature, two different types of fructose polymers (fructan) are generally found in dietary fibers; these are the fructose homopolymers levan, which is of high molecular weight and is $\beta$-(2,6)-linked, and inulin, which is of low molecular weight and is $\beta$-(2,1)-linked. The effects of levan and inulin on the intestinal physiology of rats were compared. Sprague Dawley rats were fed one of three diets for 3 weeks: a control diet, a basal diet containing 7% of levan, and a basal diet containing 7% of inulin. Cecal enlargement, together with the lowering of cecal pH, occurred in rats fed on the levan and inulin diets (p < 0.05). The levan and inulin diets resulted in a two-fold increase in the amount of short-chain fatty acids in the cecum, when compared to the control diet. The number of total microbes and of lactic acid-producing bacteria in the feces were higher in rats fed the fructan diets than those in rats fed control diet (p < 0.05). The levan diet also significantly increased the cecal $\alpha$-galactosidase activity by 3.8-fold, when compared to the control diet, indicating that levan stimulated the growth of Bifidobacteria in the cecum. These results show that the intake of levan and inulin stimulated the growth of lactic acid-producing bacteria in the cecum and thereby improved intestinal conditions in rats. (Korean J Nutrition 35(9) : 912~918,2002)

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.4
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• pp.533-537
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• 2004

The experiment was conducted to test the hypothesis that supplementing diets of lactating first parity sows with a mixture of carbohydrases (CS) improves lactation performance and second parity reproductive performance. The CS used in this study contained 7 units/g of $\alpha$-1,6-galactosidase, 22 units/g of $\beta$-1,4-mannanase, $\beta$-1,4-mannosidase and trace amounts of other enzymes. Twenty primiparous sows (Newsham Hybrid) were allotted to either the control group (no CS supplement) or the CS group (0.1% CS supplement) and fed the experimental diets during 21 d lactation period. Sows and nursing pigs were weighed at birth and weekly until weaning. Days of weaning-to-estrus were recorded. Sows had free access to feed and water. Feed intake of sows was measured daily. During the second parity gestation and lactation, all the sows were fed the same gestation and lactation diets and their reproductive performance was measured. During the second parity, there were 14 sows (7 sows per group) remained productive. For the first lactation, maternal body weight loss of the CS group was smaller (p<0.05) than that of the control group. There was no difference in litter weight gain between two groups. Voluntary feed intake of sows did not differ between the two groups. Days of weaning-to-estrus of the CS group were smaller (p<0.05) than those of the control group. In the second parity, there was no difference in the reproductive performance between the two groups. In conclusion, supplementing CS in the diet of lactating sows during the first parity decreased body weight loss and days of weaning-to-estrus of sows. However, these effects of the CS supplementation in the first parity were not successfully carried over to the second parity.

• Korean Journal of Organic Agriculture
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• v.9 no.2
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• pp.55-67
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• 2001

Exogenous enzymes which, for the purpose of this paper, include phytase, $\beta$-glucanase, pentosanase and $\alpha$-galactosidase, are now extensively used throughout the world as aditives in swine diets. The chemical effects of these enzymes are well understand. However, the manner in which their benefits to the swine are brought about is still under debate. Phytase was to increase the availability of plant phytate phosphorus, which reduces phosphorus pollution and allows reductions in the amount of inorganic phosphate used. Also, enzymes have been discovered which have the potential to break down deleterious compounds commonly found in swine rations such as $\beta$-glucanase contained in barley and oats and the soluble pentosans found in rye and wheat thus increasing the digestibility of these non-starch polysaccharides. Future research in these area will allow for more efficient use of the current enzymes, development of more efficient future products and development of more thermotolerant enzymes.

• Journal of the East Asian Society of Dietary Life
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• v.14 no.4
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• pp.363-369
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• 2004

Three different doengjang(fermented soybean paste) were prepared by using B. subtilis SS103 with high angiotensin converting enzyme(ACE) inhibitory activity, Asp. oryzae and mixed culture of two organisms, respectively. Quality characteristics and the inhibitory activity were compared. $\beta$-Amylase activity rapidly increased during 30 days of fermentation, but lower activity was found in doenjang prepared with B. subtilis. However, $\alpha$-Galactosidase activity decreased for 15 days of fermentation and little activity was observed after that fermentation period. Protease production tended to increase during the fermentation. High protease activity was found only in doengjang prepared with Asp. oryzae, but the doengjang prepared with B. subtilis SS103 showed very low activity. Total isoflavone content at 90 day fermentation was the highest in doengjang prepared with mixture of Asp. oryzae and B. subtilis. ACE inhibitory activity increased during the fermentation period. The highest activity was found in doengjang made with mixture of Asp. oryzae and B. subtilis. Sensory evaluation on doengjang itself and its soup indicated that doengjang made with mixture of Asp. oryzae and B. subtilis showed higher acceptability on taste, flavor and color than those prepared with only Asp. oryzae or B. subtilis, respectively.

• Molecules and Cells
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• v.28 no.5
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• pp.489-494
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• 2009

We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

• Mass Spectrometry Letters
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• v.5 no.3
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• pp.63-69
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• 2014

In this study, we present a new isotope-coded carbamidomethylation (iCCM)-based quantitative proteomics, as a complementary strategy for conventional isotope labeling strategies, with providing the simplicity, ease of use, and robustness. In iCCM-based quantification, two proteome samples can be separately isotope-labeled by means of covalently reaction of all cysteinyl residues in proteins with iodoacetamide (IAA) and its isotope (IAA-$^{13}C_2$, $D_2$), denoted as CM and iCCM, respectively, leading to a mass shift of all cysteinyl residues to be + 4 Da. To evaluate iCCM-based isotope labeling in proteomic quantification, 6 protein standards (i.e., bovine serum albumin, serotransferrin, lysozyme, beta-lactoglobulin, beta-galactosidase, and alpha-lactalbumin) isotopically labeled with IAA and its isotope, mixed equally, and followed by proteolytic digestion. The resulting CM-/iCCM-labeled peptide mixtures were analyzed using a nLC-ESI-FT orbitrap-MS/MS. From our experimental results, we found that the efficiency of iCCM-based quantification is more superior to that of mTRAQ, as a conventional nonisobaric labeling method, in which both of a number of identified peptides from 6 protein standards and the less quantitative variations in the relative abundance ratios of heavy-/light-labeled corresponding peptide pairs. Finally, we applied the developed iCCM-based quantitative method to lung cancer serum proteome in order to evaluate the potential in biomarker discovery study.