• The Korean Journal of Mycology
• /
• v.17 no.3
• /
• pp.149-153
• /
• 1989

One strain of Aspergillus niger K-25 producing an acid-stable ${\alpha}-amylase$ was isolated from the soil. The optimum culture conditions were investigated. The production of the acid-stable ${\alpha}-amylase$ was enhanced when the strain was incubated in a medium containing soluble starch 3.5%, peptone 2%, $KH_2PO_4$ 0.5%, $MaSO_4{\cdot}7H_2O$ 0.25% and $FeCI_3$ 1.0% at pH 3 for 7 days. However, higher activity of acid-stable ${\alpha}-amylase$ was demonstrated on wheat bran culture. Amylase production was doubled when A. niger K-25 was incubated on the wheat bran supplemented with fumaric acid buffer (pH 3).

• Applied Biological Chemistry
• /
• v.21 no.2
• /
• pp.103-108
• /
• 1978

Acid-stable ${\alpha}-amylase$ was partially purified from Paecilomyces subglobosum by Sephadex G-150 gel filtration. About 7.7-fold purification was obtained and the partially purified preparation has 5.0 U of ${\alpha}-amylase$ activity per mg of protein. Using this partially purified ${\alpha}-amylase$, general properties were studied and it showed the maximal activities at the conditions of pH 4.0 and $38^{\circ}C$. High stability of the acid-stable ${\alpha}-amylase$ in acidic condition was observed, whereas thermal stability was similar to the conventional ${\alpha}-amylase$.

• The Korean Journal of Mycology
• /
• v.17 no.3
• /
• pp.145-148
• /
• 1989

An acid-stable ${\alpha}-amylase$ produced by Aspergillus niger K-25 strain was purified by fractional precipitation with ammonium sulfate, ethacridine and acetone. The final preparation was homogeneous in cellulose acetate electrophoresis. The enzyme retained 91 % of its oringinal activity at pH 3.0, 8.7% at pH 2.4. The optimum pH of the enzyme was around pH 4. The purified-enzyme with optimum temperature of $40^{\circ}C$ was more heat-stable than the commercial product. The enzyme retained 80% of its original activity when heated to $60^{\circ}C$ for 30 minutes while the commercial amylase lost its acitivity completely within 30 minutes at $50^{\circ}C$.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.6
• /
• pp.828-836
• /
• 2015

Based on its α-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid α-amylase production was screened. The B-5 strain was identified as Bacillus amyloliquefaciens through morphological, physiological, and biochemical characteristics analysis, as well as 16S rDNA phylogenetic analysis. Its α-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant α-amylase AMY-Ba showed the optimal pH of 5.0, and was stable at a pH range of 4.0-6.0. When hydrolyzing soluble starch, amylose, and amylopectin, AMY-Ba released glucose and maltose as major end products. The α-amylase AMY-Ba in this work was different from the well-investigated J01542-type α-amylase which also came from B. amyloliquefaciens. AMY-Ba exhibited notable adsorption and hydrolysis ability towards various raw starches. Structure analysis of AMY-Ba suggested the presence of a new starch-binding domain at its C-terminal region.

• Proceedings of the Microbiological Society of Korea Conference
• /
• 1991.04a
• /
• pp.225-236
• /
• 1991

The genes encoding the thermostable $\alpha$-amylase and maltogenic amylase from Bacillus lichenciformis were cloned and expressed in E. coli. The recombinant plasmid pTA322 was found to contain a 3.1kb EcoRI genomic DNA fragment of the thermostable $\alpha$-amylase. The cloned $\alpha$-amylase was compared with the B. licheniformis native $\alpha$-amylase. Both $\alpha$-amylase have the same optimal temperature of $70^{\circ}C$ and are stable in the pH range of 6 and 9. The complete nucleotide sequences of the thermostable $\alpha$-amylase gene were determined. It was composed of one open reading rame of 1,536 bp. Start and stop codons are ATG and TAG. From the amino acid sequence deduced from the nucleotide sequence, the cloned thermostable $\alpha$-amylase is composed of 483 amino acid residues and its molecular weight is 55,200 daltons. The content of guanine and cytosine is $47.46mol\%$ and that of third base codon was $53_41mol\%$. The recombinant plasmid, pIJ322 encoding the maltogenic amylase contains a 3.5kb EcoRI-BamHI genomic DNA fragment. The optimal reaction temperature and pH of the maltogenci amylase were $50^{\circ}C$ and 7, respectively. The maltogenic amylase was capable of hydrolysing pullulan, starch and cyclodextrin to produce maltose from starch and panose from pullulan. The maltogenic amylase also showed the transferring activity. The maltogenic amylase gene is composed of one open reading frame of 1,734bp. Start and stop codons are ATG and ATG. At 2bp upstream from start codon, the nucleotide sequence AAAGGGGGAA seems to be the ribosome-binding site(RBS, Shine-Dalgarno sequence). A putative promoter(-35 and-10 regions) was found to be GTTAACA and TGATAAT. From deduced amino acid sequence from the nucleotide srquence, this enzyme was comosed of 578 amino acid residues and its molecular weight was 77,233 daltons. The content of guanine and cytosine was $48.1mol\%$. The new recombinant plasmid, pTMA322 constructed by inserting the thermostable $\alpha$-amylase gene in the EcoRI site of pIJ322 to produce both the thermostable $\alpha$-amylase and the maltogenic amylase were expressed in the E. coli. The two enzymes expressed from E. coli containing pTMA322 was reacted with the $15\%$ starch slurry at $40^{\circ}C$ for 24hours. The distribution of the branched oligosaccharides produced by the single-step process was of the ratio 50 : 50 between small oligosaccharide up DP3 and large oligosaccharide above DP3.

• Korean Journal of Microbiology
• /
• v.10 no.3
• /
• pp.106-108
• /
• 1972

The enzyme was produced by Asp.oryzae SHW 131. the enzymatic properties of .alpha.-amylase are following : 1) Crystallization of .alpha.-amylase is formed of longish square. 2) The range of stable pH is 5-10 and optimum ph is 5.5. 3) It is very unstable enzyme about EDTA and protection by $Ca^{++}$ ion and best activated at $50^{\circ}C$ about temperature. 4) Asp.oryzae SHW 131 produced .alpha.-amylase with acid-protease, neutral-protease and tepid-alkalin-protease.

• Korean Journal of Microbiology
• /
• v.10 no.3
• /
• pp.105-105
• /
• 1972

The enzyme was produced by Asp.oryzae SHW 131. the enzymatic properties of .alpha.-amylase are following : 1) Crystallization of .alpha.-amylase is formed of longish square. 2) The range of stable pH is 5-10 and optimum ph is 5.5. 3) It is very unstable enzyme about EDTA and protection by $Ca^{++}$ ion and best activated at $50^{\circ}C$ about temperature. 4) Asp.oryzae SHW 131 produced .alpha.-amylase with acid-protease, neutral-protease and tepid-alkalin-protease.

• Microbiology and Biotechnology Letters
• /
• v.21 no.6
• /
• pp.582-587
• /
• 1993

The extracellular alpha-amylase was purified to homogenity from the culture filtrate of starch grown Sch. castellii CBS 2863. The purified enzyme was glycoprotein with a molecular weight of about 56 kDa. The pH and temperature optimum were 5.5 and 40C, respectively. The enzyme was fairly stable up to 40C and at acid pH range (pH 4.0-7.0). The apparent Km and Vmax of the enzyme toward starch was 1.0mg/ml and 100U/mg protein, respectively. The analysis of amino acid composition was found to be acidic protein. The amino acid sequence of N-terminal peptide consisted of Asp-Val-Ser-Ser-Ala-X-X-Thr-Arg-Ser-Glu-Ser-Ile-Tyr.

• Journal of Ginseng Research
• /
• v.14 no.1
• /
• pp.21-26
• /
• 1990

The chemical composition and stabilities of the purified ginseng invertase were investigated. The purified enzyme was found to be a glycoprotein composed of 80.2% protein and 19.7% total sugar. The protein component of the enzyme was composed of acidic amino acid (9.3%), basic amino acid (48.9%), nonpolar amino acid (21.4%), polar amino acid (20.4%) and 6.1% S-containing amino acid. It showed especially high contents of histidine and serine. The enzyme was inactivated almost completely by the treatment with some proteases (papain, pepsin. trypsin, pancreatin and microbial alkaline pretense) and protein denatllrants (8M urea and 6M guanidine-HC1), bolt not with glyrosidase (${\alpha}$-amylase, ${\beta}$-amylase. glcoamylese and cellullase). btonosaccharides sllch as glilrose, fructose, galactose and mannose did not exert any influence on the enzyme activity. The activity of the enzyme was inhibited by Ag+, Mn2+, Hg2+, Zn2+ and Al3+, whereas Ca2+, Mg2+, Ba2+ and Fe3+ gave rather activating effects on the enzyme activity. The enzyme was relatively stable in the VH range of VH 6 and 8, and at the temperatures below 35$^{\circ}C$.

• The Korean Journal of Food And Nutrition
• /
• v.6 no.4
• /
• pp.294-300
• /
• 1993

This study was carried out to investigate the influences of cultural conditions of koji on the production of saccharogenic amylase during rice-koji making by Aspergillus awamori var. kawachii which is now widely used as koji-mold in brewing Tikju and Yakju in Korea. The optimum cultural temperature for the production of saccharogenic amylase by this mold was 36$^{\circ}C$, and at this temperature it needed 40 hours of cultivation for maximum production of this enzyme. It was favorable for high production of both organic acid and saccharogenic amylase to shift the cultural temperature form initial 36$^{\circ}C$ to 32$^{\circ}C$ after 20~25 hours of cultivation. The production of saccharogenic amylase was low when the water content of steamed rice was below 35%, but its production was high at 40~60% of water content. When the quantity of conidial inoculation was too small, the production of saccharogenic amylase was low in initial phase, but it was retrived after 40 hours of cultivation. When koji-thickness was over 3cm, the production of saccharogenic amylase was markedly restricted. The saccharogenic amylase of this koji was stable at pH 2~7, and showed high activity at pH 2~5.