• Title/Summary/Keyword: α-Amylase

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Analysis of Secretion Behavior of Human Lysozyme from Recombinant Saccharomyces cerevisiae

  • MARTEN, MARK R.;NAM SOO HAN;JIN BYUNG PARK;JIN-HO SEO
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.576-581
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    • 1999
  • Effects of signal sequences, protein sizes and dissolved oxygen on the secretion of human lysozyme from a recombinant yeast were experimentally characterized. The systems consisted of Saccharomyces cerevisiae host SEY2102 that was transformed with two different plasmids. These plasmids were identical with an exception to the plasmid pMC614, which contained the native yeast MFα1 sequence and the plasmid pMC632 with the non-native rat α-amylase signal sequence. The expression of human lysozyme was controlled by the ADHI promoter. The native yeast MFαl signal sequence was more efficient than the non-native rat α-amylase signal sequence in directing the secretion of human lysozyme. Lysozyme secreted with the α-amylase signal was retained inside the cells and released to the medium very slowly, thereby causing a lower cell growth rate and a decreased product secretion rate. Lysozyme was secreted more efficiently than invertase, which is an order of magnitude bigger in molecular size compared to lysozyme, which was under the direction of the MFαl signal sequence, suggesting that protein sizes may affect the secretion efficiency. When expressed in anaerobic conditions in the medium where the ADHI promoter was derepressed, the amount of lysozyme secreted was about twice higher than that of the aerobic culture. However, the secretion rates were identical. This result showed that the dissolved oxygen level may affect the efficiency of protein secretion only, and not the secretion rate of the product protein.

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Purification of ${\alpha}-Amylase$ Inhibitor from Black Bean in Korea (한국산 검정콩 ${\alpha}-Amylase$ 저해물질의 분리 및 정제)

  • Moon, Ju-Seok;Bae, Young-Il;Shim, Ki-Hwan
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.762-767
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    • 1995
  • The ${\alpha}-Amylase$ inhibitor from black bean(Phaseolus vulgaris) was purified to homogeneity using 70% saturated ammonium sulfate, DEAF-cellulose, Concanavalin-A sepharose chromatography and gel filtration with Superose 6. The purified α-amylase inhibitor showed a single band of 25 KD in molecular weight on the SDS-PAGE. The specific activity of the inhibitor was 544.0 units/mg and the purity was enhanced about 18-fold. The amino acids of ${\alpha}-Amylase$ inhibitor from black bean was mainly glutamic acid, aspartic acid and lysine. The inhibitor was glycoproteins and its carbohydrate contents was 3.2%.

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Quality characteristics and biological activities of Rosa multiflora Thunberg fruit through lactic acid bacteria fermentation (찔레나무 열매 젖산 발효물의 품질특성 및 생리활성)

  • EunYoung Yang;MyungHyun Kim;YoungSil Han
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.375-381
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    • 2022
  • The purpose of this study was to evaluate the quality characteristics and biological activities of Rosa multiflora Thunberg fruit extracts fermented with Lactobacillus plantarum based on fermentation period of 0, 24, 48, and 72 h. The study showed the pH of Rosa multiflora Thunberg fruit fermentation extracts have decreased as fermentation time increased, but the sugar content remained the same. The total acid content increased as the fermentation time increased. The viable cell count was at highest at 24 h (8.59 log CFU/mL) of fermentation, and the viable cell count decreased as the fermentation time increased. The total polyphenol content (14.85 mg GAE/g), total flavonoid content (6.74 mg RE/g), 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, ABTS+ radical scavenging activity, reducing power, α-glucosidase and α-amylase inhibitory activity were highest at 24 h of fermentation. Therefore, the study proved fermentation of Rosa multiflora Thunberg fruit with lactic acid increases physiological activity compared to nonfermented Rosa multiflora Thunberg fruit. Also the 24 h of fermentation had the highest activity, confirming the possibility of future use as a functional food material.

Inhibitory Effects of Sargassum thunbergii Ethanol Extract against α-amylase (지충이 에탄올 추출물의 α-amylase 저해활성)

  • Lee, So-Jeong;Song, Eu-Jin;Kim, Koth-Bong-Woo-Ri;Lee, Chung-Jo;Jung, Ji-Yeon;Kwak, Ji-Hee;Choi, Moon-Kyoung;Kim, Min-Ji;Kim, Tae-Wan;Ahn, Dong-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.6
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    • pp.648-653
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    • 2010
  • This study was performed to investigate the inhibitory activity of Sargassum thunbergii (ST) against ${\alpha}$-amylase and elucidate the availability of ST extract as a functional food agent. To test the inhibitory activity of ST against ${\alpha}$-amylase, porcine pancreatic ${\alpha}$-amylase and potato starch were used as substrates. It was revealed that ST crude ethanol extracts have high ${\alpha}$-amylase inhibitory activity. Subsequently, ST crude ethanol extract was separated into five partition layers by solvent extraction: n-hexane, chloroform, ethyl acetate, butanol, and water. Chloroform and n-hexane fractions showed higher inhibitory activities than did acarbose (positive control). To confirm the changes in enzyme inhibitory activity by physical treatments, ST crude ethanol extract was subjected to heat, pH, and ${\gamma}$-irradiation treatments. In all heat treatments with the exception of one ($121^{\circ}C$, 15 min), the inhibitory activity was increased compared with the untreated group. With regard to pH stability, ST extract showed no significant changes at pH 4.6, but somewhat decreased inhibitory activity was revealed at pH 2, 8, and 10. On the other hand, ST ethanol extract was stable under ${\gamma}$-irradiation under all conditions (3.20 kGy). In summary, ST ethanol extract can be used in the food industry as a natural ${\alpha}$-amylase inhibitor.

Influence of Rumen Escape Starch on α-Amylase Activity in Pancreatic Tissue and Small Intestinal Digesta of Lambs

  • Xu, M.;Yao, J.H.;Wang, Y.H.;Wang, F.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.12
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    • pp.1749-1754
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    • 2006
  • Two slaughter experiments were conducted to determine the effects of rumen escape starch, by altering dietary starch concentration and corn particle size, on ${\alpha}$-amylase activity in the pancreas and the small intestinal digesta of lambs. In experiment 1, 18 wether lambs (28.5${\pm}$1.6 kg) were fed low, medium or high starch diets for 35 d and slaughtered. Dietary starch concentrations linearly increased rumen escape starch (p<0.05). Pancreatic ${\alpha}$-amylase activity was lower (p<0.05) in lambs fed the low starch diet. When expressed per gram of digesta, ${\alpha}$-amylase activity was lower in lambs fed the low starch diet. However, expressed as total activity, ${\alpha}$-amylase in the digesta was greater in lambs fed the medium starch diet. In experiment 2, 12 wether lambs (23.5${\pm}$0.3 kg) were fed diets with finely cracked corn, coarsely cracked corn and whole corn. These dietary treatments continued for 35 d before tissue collection. Rumen escape starch increased with increasing corn particle size (p<0.05). ${\alpha}$-amylase activity in the pancreas and the small intestinal digesta was significantly greater (p<0.05) in lambs fed the coarsely cracked corn. These data suggest that increasing rumen escape starch results in a quadratic increase in total ${\alpha}$-amylase activity in the pancreas and the small intestinal digesta. Maximum ${\alpha}$-amylase activity is reached when rumen escape starch is about 100-120 g/d in 25-30 kg lambs.

Immobilization of α-amylase from Exiguobacterium sp. DAU5 on Chitosan and Chitosan-carbon Bead: Its Properties

  • Fang, Shujun;Chang, Jie;Lee, Yong-Suk;Hwang, Eun-Jung;Heo, Jae Bok;Choi, Yong-Lark
    • Journal of Applied Biological Chemistry
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    • v.59 no.1
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    • pp.75-81
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    • 2016
  • Glutaraldehyde was used as a cross-linking agent for immobilization of purified ${\alpha}$-amylase from Exiguobacterium sp. DAU5. Befitting concentration of glutaradehyde and cross-linking time is the key to preparation of cross-linking chitosan beads. Based on optimized immobilization condition for ${\alpha}$-amylase, an overall yield of 56% with specific activity of 2,240 U/g on chitosan beads and 58% with specific activity of 2,320 U/g on chitosan-carbon beads was obtained. The optimal temperature and pH of each immobilized enzyme activity were $50^{\circ}C$ and 50 mM glycine-NaOH buffer pH 8.5, respectively. Those retained more than 75 and 90% of its maximal enzyme activity at pH 7.0-9.5 and after incubation at $50^{\circ}C$ for 1 h, respectively. In addition, the immobilization product showed higher organic-solvent tolerance than free enzymes. The mode of hydrolyzing soluble starch revealed that the ${\alpha}$-amylase possessed high hydrolyzing activity. These results indicate that chitosan is good support and has broad application prospects of enzyme immobilization.

Effects of Heat and pH Treatments on α-Amylase Inhibitory Activity of Ecklonia cava Ethanol Extract (감태(Ecklonia cava) 에탄올 추출물의 α-Amylase 저해활성에 미치는 열 및 pH의 영향)

  • Kim, Dong-Hyun;Jung, Ji-Yeon;Kim, Koth-Bong-Woo-Ri;Lee, Chung-Jo;Kwak, Ji-Hee;Kim, Min-Ji;SunWoo, Chan;Kim, Hyun-Jee;Jung, Seul-A;Kim, Tae-Wan;Cho, Young-Je;Ahn, Dong-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.6
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    • pp.791-795
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    • 2011
  • This study examined the inhibitory activity of Ecklonia cava (EC) against ${\alpha}$-amylase to evaluate the availability of EC extract as a functional food agent. To verify the inhibitory activity of EC against porcine pancreatic ${\alpha}$-amylase, potato starch was used as a substrate. This analysis revealed that EC ethanol extract exhibited high ${\alpha}$-amylase inhibitory activity. For potential application within the food industry, the stability of the activity of EC ethanol extract under various heat and pH conditions was examined. The ${\alpha}$-amylase inhibitory activity of EC ethanol extract was not affected by the heat and pH treatment conditions used in this study. These results suggest that EC has the potential for development as a functional food agent.

Gelidium amansii Extract, a Potent α-glucosidase and α-amylase Inhibitor, Alleviates Postprandial Hyperglycemia in Diabetic Mice (당뇨 마우스에서 우뭇가사리(Gelidium amansii)의 식후 고혈당 완화 효과)

  • Park, Jae-Eun;Kim, Jung-Min;Han, Ji-Sook
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1052-1058
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    • 2017
  • Gelidium amansii shows antioxidant and anti-obesity effects; however, the effect on postprandial blood glucose levels is not known. The objective of the present study was to investigate the inhibitory effect of Gelidium amansii extract (GAE) on carbohydrate-digesting enzymes and its ability to alleviate postprandial hyperglycemia in streptozotocin (STZ)-induced diabetic mice. Gelidium amansii was extracted with 80% ethanol and concentrated for use in this study. The ${\alpha}-glucosidase$ and ${\alpha}-amylase$ inhibition assays were performed using the colorimetric method. ICR normal and STZ-induced diabetic mice were orally administered GAE (300 mg/kg body weight) or acarbose (100 mg/kg body weight) alone or soluble starch (2 g/kg body weight). Blood samples were taken from the tail vein at 0, 30, 60 and 120 min. Our results indicated that GAE markedly inhibited ${\alpha}-glucosidase$ and ${\alpha}-amylase$ activities with $IC_{50}$ values of $0.099{\pm}0.009mg/ml$ and $0.178{\pm}0.038mg/ml$, respectively, and was a more effective inhibitor than acarbose, the positive control. Further, the postprandial blood glucose levels of STZ-induced diabetic mice in the GAE-administered group were significantly lower than those of control group mice (p<0.05). Moreover, the area under the curves (AUC) significantly decreased with GAE administration in STZ-induced diabetic mice (p<0.05). These results indicate that GAE may be effective in decreasing postprandial blood glucose levels by inhibiting carbohydrate-digesting enzymes such as ${\alpha}-amylase$ and ${\alpha}-glucosidase$. Therefore, GAE could be used as a potential functional food for alleviating postprandial hyperglycemia.

Preparation of Red Ginseng Extract Rich in Acidic Polysaccharide from Red Tail Ginseng Marc Produced After Extraction with 70% Ethyl Alcohol (홍미삼 알콜 추출박을 이용한 산성다당체 다량 함유 홍삼 엑기스 제조)

  • 도재호;이종원
    • Journal of Ginseng Research
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    • v.20 no.1
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    • pp.60-64
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    • 1996
  • In this study, we investigated the appropriate conditions to extract acidic polysaccharide and to prepare red ginseng extract being rich in acidic polysaccharide from red tail ginseng marc produced after manufacturing alcoholic extract from red tail ginseng. Amount of acidic polysaccharide in red tail ginseng marc was about 11%. The best condition for the extraction of acidic polysaccharide from the marc was using of 3~5 mg of $\alpha$-amylase/g residue/15 ml of distilled water, and the amount of acidic polysaccharide in water extract of the residue treated with $\alpha$-amylase was about 27%. So, it is possible to manufacture red ginseng extract being rich in acidic polysaccharide using water extract of red tail ginseng alcoholic residue as extraction solvent. From the above results, we suggest that red tail ginseng residue produced by manufacturing alcoholic extract of red tail ginseng has high potencies in the utilization of waste material.

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Bioactive Properties of Novel Probiotic Lactococcus lactis Fermented Camel Sausages: Cytotoxicity, Angiotensin Converting Enzyme Inhibition, Antioxidant Capacity, and Antidiabetic Activity

  • Ayyash, Mutamed;Olaimat, Amin;Al-Nabulsi, Anas;Liu, Shao-Quan
    • Food Science of Animal Resources
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    • v.40 no.2
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    • pp.155-171
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    • 2020
  • Fermented products, including sausages, provide several health benefits, particularly when probiotics are used in the fermentation process. This study aimed to examine the cytotoxicity (against Caco-2 and MCF-7 cell lines), antihypertensive activity via angiotensin-converting enzyme (ACE) inhibition, antioxidant capacity, antidiabetic activity via α-amylase and α-glucosidase inhibition, proteolysis rate, and oxidative degradation of fermented camel and beef sausages in vitro by the novel probiotic Lactococcus lactis KX881782 isolated from camel milk. Moreover, camel and beef sausages fermented with commercial starter culture alone were compared to those fermented with commercial starter culture combined with L. lactis. The degree of hydrolysis, antioxidant capacity, cytotoxicity against Caco-2 and MCF-7, α-amylase, α-glucosidase, and ACE inhibitory activities were higher (p<0.05) in fermented camel sausages than beef sausages. In contrast, the water and lipid peroxidation activity were lower (p<0.05) in camel sausages than beef sausages. L. lactis enhanced the health benefits of the fermented camel sausages. These results suggest that camel sausage fermented with the novel probiotic L. lactis KX881782 could be a promising functional food that relatively provides several health benefits to consumers compared with fermented beef sausage.