• Biotechnology and Bioprocess Engineering:BBE
• /
• v.8 no.2
• /
• pp.106-111
• /
• 2003

A novel carbohydrolase, which is a DXAMase, containing both dextranase and amylase equivalent activities, was purified from Lipomyces starkeyi KSM22. The purified DXAMase was also found to hydrolyze cellobiose, gentiobiose, trehalose and melezitose, while disproportionation reactions were exhibited with various di- and tri-saccharides, such as maltose, isomaltose, gentiobiose, kojibiose, sophorose, panose, maltotriose, and isomaltotriose with various kinds of oligosaccharides produced as acceptor reaction products. Furthermore, the purified DXAMase hydrolyzed raw waxy rice Starch and produced maltodextrin to the extent of 50% as a glucose equivalent.

• Microbiology and Biotechnology Letters
• /
• v.25 no.2
• /
• pp.109-114
• /
• 1997

The Oomycete Saprolegnia ferax produces an extracellular $\beta$-amylase, Maximum enzyme yield was attained after 7 days of growth in YNB starch medium (pH 6.5) at 25$\circ$C. The amylase was pu- rified 24-fold by ultrafitration, HPLC DEAE column and HPLC gel filtration. The purfied enzyme was a monomeric glycoprotein with a molecular weight of about 44,000 dalton. The pH and temperature optima were 6.5 and 50$\circ$C, respectively. The enzyme was fairly stable up to 50$\circ$C and at acidic pH region (pH 4.0-7.0). The apparent Km and Vmax values of the enzyme against soluble starch were 0.77 mg/ml and 2,174 $\mu$moles/mg protein, respectively. Amino acid analysis indicated that the enzyme was enriched in alanine, glycine, leucine and acidic amino acid. Starch hydrolysis with the enzyme released maltose but not glucose, whereas maltotriose, Schardinger dextrin ($\alpha$-cyclodextrin) and pullulan were not hydrolysed by the enzyme. The enzyme was inhibited by Schardinger dextrin, p-chloromercuribenzoate(PCMB), CU$^{2+}$' and Hg$^{2+}$. Inhibition of the enzyme by PCMB could be reversed by the addition of cysteine and mercaptoethanol.

• Food Science and Biotechnology
• /
• v.14 no.5
• /
• pp.681-684
• /
• 2005

A maltopentaose-producing amylase (G5-amylase) from Bacillus megaterium KSM B-404 was applied to retard bread retrogradation. Retrogradation rates were determined by differential scanning calorimetry. Gel permeation chromatography determined changes in maltooligosaccharide composition and the molecular weight profiles of carbohydrate tractions. The baking process produced maltopentaose and maltotriose by the hydrolysis of starch molecules into small units. Amylose and amylopectin degradation as well as maltooligosaccharides produced by the enzyme were likely responsible for retarding starch retrogradation. Overall, addition of G5-amylase reduced the starch retrogradation rate, and was as effective as Novamyl(R), a commercial enzyme.

• Food Science and Biotechnology
• /
• v.14 no.1
• /
• pp.32-38
• /
• 2005

Effects of acid and ${\alpha}$-amylase on resistant starches including retrograded RS3 and cross-linked RS4 prepared from normal maize starch were investigated. Acid and ${\alpha}$-amylase hydrolytic patterns of RS3 were similar, while those of native starch and RS4 differed. Acid hydrolysis rate of RS3 was markedly higher at initial stage, then slowly decreased up to 20 days, whereas that of RS4 increased continuously. The sizes of acid- and ${\alpha}$-amylase-treated RS3 residues decreased, but those of RS4 remained unchanged. X-ray patterns of all treated residues did not change; however, the peak intensities increased. Swelling power of RS3 increased to 150% at $95^{\circ}C$, whereas that of RS4 differed depending on the treatment condition. Swelling power of acid-treated RS4 residue increased markedly, but that of ${\alpha}$-amylase-treated one remained constant. Gel filtration chromatography profiles of untreated RS3 and RS4 residues were similar, whereas that of acid-treated RS4 residue was different from them. RS showed different hydrolytic behavior by acid and ${\alpha}$-amylase depending on the type, and susceptibility of RS3 was higher than that of RS4.

• Journal of Ginseng Research
• /
• v.28 no.3
• /
• pp.136-142
• /
• 2004

During our investigations on the relationship between the browning reaction of ginseng root and two compounds (arginyl-fructosyl-glucose and arginyl-fructose) in the model system of steaming and heat-drying processes for the preparation of red ginseng, the preheating treatment of main roots of raw ginseng at 60∼70$^{\circ}C$ prior to the steaming and heat-drying processes was found to bring about the gelatinization of starch granules. The enzymatic hydrolysis of gelatinized starch to maltose, a marked formation of maltose, and the increase of both free arginine and total amino acids, resulting the acceleration of the Maillard type-browning reaction of ginseng root during the steaming and heat-drying processes, and the rise of brown color intensity of red ginseng. These results show that the preheating treatment may be effective for the decrease of inside white of red ginseng.

• Applied Biological Chemistry
• /
• v.35 no.5
• /
• pp.389-394
• /
• 1992

Effect of dry-heat treatment $(130{\sim}220^{\circ}C)$ on properties of corn starch was investigated. No significant changes of surface color and swelling power were observed. Blue value, water-binding capacity, solubility and degree of hydrolysis by acid or glucoamylase were increased; however, intrinsic viscosity and crystallinity were decreased as heating temperature was raised. Noticeable changes occurred between $170{\sim}190^{\circ}C$.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.8
• /
• pp.1401-1407
• /
• 2008

The roles of conserved amino acid residues (Va1329-Ala330-Asn331-Glu332), constituting an extra sugar-binding space (ESBS) of Thermus maltogenic amylase (ThMA), were investigated by combinatorial saturation mutagenesis. Various ThMA mutants were firstly screened on the basis of starch hydrolyzing activity and their enzymatic properties were characterized in detail. Most of the ThMA variants showed remarkable decreases in their hydrolyzing activity, but their specificity against various substrates could be altered by mutagenesis. Unexpectedly, mutant H-16 (Gly-Leu-Val-Tyr) showed almost identical hydrolyzing and transglycosylation activities to wild type, whereas K-33 (Ser-Gly-Asp-Glu) showed an extremely low transglycosylation activity. Interestingly, K-33 produced glucose, maltose, and acarviosine from acarbose, whereas ThMA hydrolyzed acarbose to only glucose and acarviosine-glucose. These results propose that the substrate specificity, hydrolysis pattern, and transglycosylation activity of ThMA can be modulated by combinatorial mutations near the ESBS.

• Journal of Applied Biological Chemistry
• /
• v.49 no.4
• /
• pp.135-139
• /
• 2006

A alkalophilic strain, Bacillus licheniformis MH31 producing an alkaline protease was isolated from mine soil of Boryeong in Korea. Production of a high level of alkaline protease was achieved 42 h after incubation when the bacterium was grown at pH 9.0 and $35^{\circ}C$ in Horikoshi medium supplemented with 0.5%(w/v) starch and 1%(w/v) skim milk as carbon and nitrogen source, respectively. The molecular weight of partially purified enzyme was estimated to be 30 kDa by SDS-PAGE and its optimum pH was pH 10. The enzyme showed optimum temperature at $50^{\circ}C$, and was stable up to $60^{\circ}C$ after 1 h incubation. The protease was strongly inhibited by 1 mM of PMSF which was known well as strong inhibitor of serine proteases, but almost not inhibited by 5 mM of EDTA and 1,10-phenanthroline. When the protein hydrolysis products of 1% skim milk by partially purified protease was compared with available commercial proteases using HPLC analysis, most of hydrolysis products were detected below molecular weight of 10,000 and the hydrolysis ratio of purified enzyme was 24.8% lower than those(above 32%) of commercial proteases.

• Microbiology and Biotechnology Letters
• /
• v.14 no.5
• /
• pp.359-363
• /
• 1986

S. diastaticus hydrolysised $\alpha$-1.4 linkage of the starch and was known fermenting yeast strain, but poorly hydrolized $\alpha$-1.6 linkage of the starch. To improve the starch fermentation ability of yeast, we tried that protoplast fusion between S. diastaticus and C. tropicalis and finally two starins of fusant (FPDC42, FPDC43) were obtained. C. tropicalis well hydrolysis both $\alpha$-1.4 and $\alpha$-1.6 linkanges in the starch. The protoplast of parental auxotrophic cells were fused in the presence of 10mM CaCl$_2$ and 35% of polyethylene glycol (M. W. 4,000). The fusion frequency was 10$^{-5}$ to 10$^{-6}$. Properties of the fusants(genetic stability, assimilation of carbon sources, random spore formation, copper resistance, NaCl tolerance, DNA content, cell size and growth rate) were investigated.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.1
• /
• pp.122-126
• /
• 2004

A digestion trial in vitro was conducted to study effects of supplementation of NSP (non-starch polysaccharides) degrading enzyme (feed grade) on cell wall degradation and digestibility of nutrients in barley. The slices of barley were soaked in distilled water with or without 0.15% non-starch polysaccharides degrading enzyme. Microscopic examination of the slices showed that the endosperm cell wall of barley was completely degraded by the non-starch polysaccharides degrading enzyme. The residues and supernatant of digesta in vitro were separated by filtration with 0.1 mm nylon fabric. The residues were used for measurement of crude protein, crude fat, crude fiber, and moisture. The supernatant was used for determination of viscosity, as well as amino-nitrogen and glucose content. The results showed that compared with the control, the amino-nitrogen and glucose content of the supernatant increased by 17.58% (p<0.05) and 10.26% (p<0.05), respectively, while viscosity did not change. Enzyme supplementation increased the digestibilities of dry matter, crude protein, nitrogen-free extract, crude fat and crude fiber of barley by 18.1% (p<0.05), 20.3% (p<0.05), 16.4% (p<0.05), 26.9% (p<0.05) and 30.0% (p<0.05), respectively. The present study suggests that cell wall hydrolysis may contribute to improved nutrient digestion in vivo when non-starch polysaccharides degrading enzymes are fed to swine.