• Title/Summary/Keyword: acid-stable ${\alpha}-amylase$

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Novel $\alpha$-Glucosidase from Extreme Thermophile Thermus caldophilus GK24

  • Nashiru, Oyekanmi;Koh, Suk-Hoon;Lee, Se-Yong;Lee, Dae-Sil
    • BMB Reports
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    • v.34 no.4
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    • pp.347-354
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    • 2001
  • $\alpha$-Glucosidase of an extreme thermophile, Thermus caldophilus GK24 (TcaAG), was purified 80-fold from cells to a homogeneous state and characterized. The enzyme exhibited optimum activity at pH 6.5 and $90^{\circ}C$, and was stable from pH 6.0 to 85 and up to $90^{\circ}C$. The enzyme had a half-life of 85 minutes at $90^{\circ}C$. An analysis of the substrate specificity showed that the enzyme hydrolyzed the non-reducing terminal unit of $\alpha$-1,6-glucosidic linkages of isomaltosaccharides and panose, $\alpha$-1,3-glycosidic bond of nigerose and turanose, and $\alpha$-1,2-glycosidic bond of sucrose. The gene encoding the TcaAG was cloned, sequenced, and sequenced in E. coli. The nucleotide sequence of the gene encoded a 530 amino acid polypeptide and had a G+C content of 68.4% with a strong bias for G or C in the third position of the codons (93.6%). A sequence analysis revealed that TcaAG belonged to the $\alpha$-amylase family. We suggest that this monomeric, thermostable, and broad-acting $\alpha$-glucosidase is a departure from previously exhibited specificities. It is, therefore, a novel $\alpha$-glucosidase.

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Correlation between Nutrient Components of Grass Roughages Mainly Used in Korea (국내 주요 화본과 조사료의 영양성분 간 상관관계)

  • Kim Myeong Hwa;Seo In Jun;Kim Joon Sun;Lee Sang Rak
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.3
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    • pp.169-176
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    • 2005
  • To evaluate the possibility of predicting a specific nutrient component, which nay be difficult in analysis, from a component of nutrient relatively easy to analyze, six grass roughage sources (bermuda grass hay, klein grass hay, oat hay, orchard grass straw, tall fescue straw and timothy hay) mainly used in Korea were analyzed for the heat-stable $\alpha-amylase$ treated neutral detergent fiber (aNDF), acid detergent fiber (ADF), acid detergent lignin (ADL), neutral detergent insoluble crude protein (NDICP) and acid detergent insoluble crude protein (ADICP) as well as proximate analytical components, and correlation coefficients between nutrient components were calculated. Some chemical components appeared to have significant correlation (p<0.05) through Pearson's correlation coefficients (i.e. between crude protein (CP) and NDICP in oat hay, orchard grass straw and tall fescue, between CP and ADICP in oat hay and timothy hay, between crude fiber (CF) and aNDF in klein grass hay, tall fescue straw and timothy hay, between CF and ADF in oat hay, tall fescue straw and timothy hay, and between CF and ADL in orchard pass straw and timothy hay). It is therefore suggested that it would be worthy to predict a specific nutrient component, which may be difficult in analysis such as aNDF, ADF, ADL, NDICP and ADICP, from a component of nutrient relatively easy to analyze such as CP and CF. However, it would be needed to improve the correlation coefficients, more samples should be measured.

Characterization of Exo-dextranase from Aspergillus ustus (Aspergillus ustus의 Exo-dextranase의 특성에 관한 연구)

  • Lee, Kon-Joo;Lee, Hyung-Hoan
    • The Korean Journal of Mycology
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    • v.11 no.1
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    • pp.15-21
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    • 1983
  • Exodextranase from Aspergillus ustus was purified by chromatography and characterized by various conditions. The optimal pH of the purified dextranase was 6.5 and this enzyme was maximally activated at $40^{\circ}C$. The enzyme was stable at the temperature below $50^{\circ}C$. The enzyme was markedly inactivated by $Hg^{2+},\;Cu^{2+},\;KCN\;and\;Co^{2+},\;but\;Ba^{2+},\;Fe^{2+},$ cysteine, EDTA, and ascorbic acid enhanced the activity of the enzyme. The main products from the hydrolysis of dextran incubated with the dextranase were glucose, isomaltotriose and oligosaccharide. When dextran was incubated with the mixture of pullulanase and ${\alpha}-amylase$, it was hydrolyzed into glucose, isomaltose and oligosaccharide. Polysaccharides in the decade teeth powder were hydrolyzed by the dextranase into glucose, isomaltotriose and oligosaccharides. In the hydrolysis of the teeth powder with the mixture of dextranase, pullulanase and ${\alpha}-amylase$, were proved to be similar to the dextran hydrolysates.

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Effect of Germination Condition and Drying Methods on Physicochemical Properties of Sprouted Brown Rice (발아조건 및 건조방법이 발아현미의 이화학적 특성에 미치는 영향)

  • 김선림;손영구;손종록;허한순
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.3
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    • pp.221-228
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    • 2001
  • This study was carried out to investigate the effect of germination condition and drying temperature on growth and physicochemical properties of brown rice. Three brown rice seeds of Ilpumbyeo, Dasanbyeo and Heugjinjubyeo were stored at room temperature for six weeks to test the time-sequence germination viability. Relatively stable germination ratio was maintained until 2 weeks after storage. However, 3 weeks after storage, germination ratio of brown rice seeds started to decrease rapidly and their germination ratio was lower than 80%. For this reason, brown rice was recommended for seeding within 2 weeks after hulling. During the initial 5 days, germination ratio of 24 hours pre-soaking brown rice was higher about 2-3% than that of non-soaking brown rice. The $25^{\circ}C$ was considered as the most favorable temperature for brown rice germination, because of the high germination ratio and desirable coleoptile growth of the brown rice, and little seed rotting symptoms. The scanning electron micrographs showed the structural differences between hot-air dried and freeze dried germinated-brown rice kernel. In the freeze dried germinated-brown rice, seed coat (pericarp, tegmen and aleurone layer) was mechanically disrupted from the endosperm, and many cleavages were observed among starch storing cells and starch granules. The endosperm of freeze-dried brown rice kernels formed the sponge-like structures and showed the fragile traits. For this reason, hot-air drying is considered as more suitable method than freeze drying for germinated-brown rice. The crude protein and amylose contents were slightly changed, but there were no significant differences during the germination period. Crude fiber content was decreased, but crude Int and total amino acid contents were increased as seeding days increased. A rapid increase in $\alpha$-amylase activities of germinating brown rice was observed at S days after seeding, and $\alpha$-amylase activities were decreased from 8 days after seeding. Total free sugar contents were decreased during the germination period. There was continuous decline in the contents of sucrose and glucose until 8 days after seeding, but fructose and maltose content were gradually increased from the 5 days after seeding.

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Selection of Acid-tolerant and Hetero-fermentative Lactic Acid Bacteria Producing Non-proteinaceous Anti-bacterial Substances for Kimchi Fermentation (비단백질성 항균물질을 생산하는 김치발효용 내산성 Hetero 발효형 유산균주 선발)

  • Kim, Hye-Rim;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.119-127
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    • 2013
  • Twenty-three strains of Leuconostoc species and 45 strains of Weissella species inhibiting the growth of Lactobacillus sakei, one of the most populous lactic acid bacteria in over-ripened kimchi, were isolated from kimchi in our previous study. Among these hetero-fermentative 68 strains, Leuconostoc mesenteroides CK0128, Weissella cibaria CK0633, and W. cibaria KK0797 exhibited a relatively high survival rate in MRS medium, which was adjusted to pH 4.3 using an acid mixture consisting of acetic and lactic acids, and produced a large amount of exopolysaccharides. The culture supernatants of 3 strains were fractionated by a molecular weight cutter and lyophilized. The fractions with a molecular weight smaller than 3,000 Da showed antagonistic activity against Staphylococcus aureus and Lb. sakei. The anti-bacterial substances were very stable to heat treatments ($121^{\circ}C$, 15 min) and active at acidic conditions below pH 5. ${\alpha}$-Amylase, lipase, and proteolytic enzymes (proteinase K and pepsin) did not affect their activities. These non-proteinaceous anti-bacterial substances inhibited the growth of several food pathogens.

Polymerization of Sugar by Extrusion

  • Hwang, Jae-Kwan;Kim, Chul-Jin;Chong-Tai, Kim
    • Preventive Nutrition and Food Science
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    • v.2 no.4
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    • pp.296-300
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    • 1997
  • Glucose syrup and lactose, mixed with citric acid as a polymerizing catalyst, was processed using twin screw extruder, in which 40 of L/D(length/diameter) ratio was designed to provide sufficient retention in extruder for polymerization of sugars. The polymerization yields of glucose syrup were 36.90%, 55.44% and 77.10% at 160, 180 and 20$0^{\circ}C$, respectively, while those of lactose were 26.45%, 38.16% and 45.86% at the same temperatures. Gel permeation chromatography exhibited that the higher molecular weight fractions were increased with extrusion temperature, which also led to increasing hydrodynamic intrinsic viscosity. Both uco-oligosaccharides and lacto-oligosaccharides produced by extrusion of glucose syrup and lactose were stable for thermal treatments over a wide range of pH3.0~11.0. In addition, $\alpha$-amylase and amyloglucosidase treatment of gluco-oligosaccharides did not affect the solution viscosity, indicating the random linkage rather than $\alpha$-1, 4 linkages of glucose and thus the potential applications as a dietary fiber. In this research it was clearly observed that twin screw extrusion can be successfully utilized to produce gluco-oligosaccharides and lacto-oligosaccharides rapidly and continuously in conjunction with selective control of polymerized composition.

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Bacteriocin Produced by Pediococcus sp. in Kimchi and Its Characteristics

  • Kwon, Dae-Young;Koo, Min-Seon;Ryoo, Chae-Ran;Kang, Chang-Hoon;Min, Kyung-Hee;Kim, Wang-June
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.96-105
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    • 2002
  • A bacteriocin-producing strain identified as Pediococcus acidilactici was isolated from kimchi. The bacteriocin was identified to belong to the pediocin family and exhibited bactericidal activity against most Gram-positive bacteria as well as some Gram-negative bacteria. The bacteriocin was stable up to $80^{\circ}C with wide pH ranges (5.0-10.0). The bactericidal activity remained unchanged after treatment with nonproteolytic enzymes such as nuclease and ${\alpha}$-amylase, however, it was destroyed after treatment with protease. The bacteriocin was effectively extracted by the pH-mediated adsorption-desorption method and purified effectively by semi-preparative RP-HPLC. The molecular weight of the bacteriocin was 4,622, as determined by electrospray mass spectrometry. The amino acid sequence consisted of 44 amino acid residues with four cysteines. The high solubility and pH stability of the isolated pediocin provide definite advantages over nisin and other bacteriocins in regards to its potential applications.

Bactericidal Effect of Bacteriocin of Lactobacillus plantarum K11 Isolated from Dongchimi on Escherichia coli O157

  • Lim, Sung-Mee;Im, Dong-Soon
    • Journal of Food Hygiene and Safety
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    • v.22 no.3
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    • pp.151-158
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    • 2007
  • Among 68 strains of lactic acid bacteria (LAB) isolated from Dongchimi, a strain K11 was selected due to its bactericidal activity against Escherichia coli O157 The strain K11 was identified as Lactobacillus plantarum, based on physiological and biochemical characteristics. In the late exponential phase, La. plantarum K11 showed maximum bacteriocin activity (12,800 BU/mL) and maintained until the early stationary phase. The bacteriocin activity was completely inactivated by all the proteolytic enzymes such as pepsin, protease, proteinase K, papain, chymotrypsin, and trypsin, but the activity was not affected by catalase, a-amylase, lysozyme, and lipase, suggesting proteinaceous nature of the bacteriocin. Additionally, this activity was not affected in the pH range from 3.0 to 9.0 and under storage conditions like 30 days at -20,4, or $25^{\circ}C$. Although the bacteriocin activity was absolutely lost after 15 min treatment at 121, it was relatively stable at $70^{\circ}C$ for 60 min or $100^{\circ}C$ for 30 min. The activity was disappeared by treatment with acetone, benzene, ethanol, or methanol, but it was not affected by treatment with chloroform or hexane. The antibacterial activity of the bacteriocin was good against some LAB including Lactobacillus spp., Enterococcus spp., and Streptococcus spp., but not against food-borne pathogens such as Bacillus spp., Listeria spp., and Staphylococcus spp. as well as yeasts and molds. Especially, some intestinal bacteria such as Enterobacter aerogenes and E. coli were significantly affected by the bacteriocin of La, plantarum K11. Furthermore, the addition of 640 BU/mL resulted in the complete clearance of E. coli O157 after 10 hr.

Chayacterization of Bacillus polyfermenticus SCD as a Probiotic. (Bacillus polyfermenticus SCD의 생균제로서의 특성)

  • 전경동;김혜진;이광호;백현동;강재선
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.359-366
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    • 2002
  • Bacillus polyfermenticus SCD which is commonly called as Bisroot strain is being used for functional foods through the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine in humans. The cells of B. polyfermenticus SCD were treated for 4h in artificial gastric juice (pH 2.0,3.0) and bile acid. Final viability of the strain in artificial gastric Juice (pH 2.0, 3.0) is reached to 62.8% and 81.2% respectively B. polyfermenticus SCD is resistant to antibiotics such as streptomycin, rifampicin, nystatin and ampicilin. B. polyfermenticus SCD is well known supplies the nutrients by synthesizing vitamin $B_1$, $B_2$, C and K. B. polyfermenticus SCD produces various digestive enzymes and the enzymes enable to completely digest diets in our body. Above all, $\alpha$-amylase and pretense activities are very higher than B. subtilis KCTC 1020, about two fold and twenty five fold respectively. B. polyfermenticus SCD is very stable during long-term storage period in phosphate buffers of wide-range pH, solutions of various concentrations of sodium chloride, 5% glucose solution and water.