• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.417-422
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• 1997

Recombinant Bacillus subtilis LKS88[pASA240] containing the amylase gene from Streptomyces albus KSM-35 was exploited in fed-batch cultivation for mass production of maltotetraose-producing amylase. The effects of dissolved oxygen, additional organic nutrients (peptone and yeast extract) and mixed carbon sources (glucose plus soluble starch) on amylase production were examined in fed-batch operations in an effort to determine the optimum conditions for a maximum amylase productivity. Under the optimum conditions, maximum amylase activity was about 4.2 times higher than that obtained in batch cultivations, indicating that mass production of maltotetraose-producing amylase could be accomplished in fed-batch cultivation of the recombinant B. subtilis strain.

• Korean Journal of Food Science and Technology
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• v.26 no.5
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• pp.633-637
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• 1994

A bacterial strain KSM-35 producing maltotetraose forming amylase was isolated from compost and identified as Streptomyces based on its morphological, cultural, and physiological characteristics. The amylase from Streptomyces sp. KSM-35 culture filtrate was purified by ammonium sulfate precipitation, followed by the liquid chromatographic procedures using DEAE-Toyo pearl and sephadex G-100 with 27.1% activity recovery. The molecular weight of the enzyme was estimated to be 50,000 and the isoelectric point 4.3. The main product by the amylase from soluble starch was maltotetraose which accounted for 56% of all the oligosaccharides detected after 26 hrs of reaction. Maltose (20%o) and maltotriose (16%) were the next important byproducts while glucose and maltopentaose were detected as traces.

• KSBB Journal
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• v.16 no.3
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• pp.246-252
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• 2001

We isolated a bacterium that produces an extracellular maltopentaose(G5)-forming amylase from amylose and soluble starch. The bacterium was identified and assigned as a Bacillus sp. AIR-5. The amylase did not hydrolyze maltose, maltotriose, maltotetraose or maltopentaose. Optimum medium composition for maltopentaose production in flask culture was 2%(w/v) soluble starch, 0.4%(w/v) tryptone, 0.5%(w/v) NaCl, 0.5%(w/v) K$_2$HPO$_4$, and 3 mM CaCl$_2$at pH 8.0, 28$^{\circ}C$. The highest yield for maltopentaose production in this condition was 6.45 g/L and was 32.55% of theoretical yield.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.573-579
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• 1995

A Bacillus strain capable of producing an extracellular malto-oligosaccharides forming $\alpha $-amylase was isolated from soil and designated as Bacillus sp. SUH4-2. The enzyme was purified by ammonium sulfate fractionation, DEAE-Toyopearl and Mono-Q HR 5/5 column chromatographies using a FPLC system. The specific activity of the enzyme was increased by 16.1-fold and the yield was 13.5%. The optimum temperature for the activity of $\alpha $-amylase was 60-65$\circ$C and more than 50% of initial activity was retained after the enzyme was incubated at 60$\circ$C for 40 min. The enzyme was stable over a broad pH range of 5.0-8.0 and the optimum pH was 5.0-6.0. The molecular weight of the enzyme was determined to be about 63.6 kD and isoelectric point was around 5.8. The enzyme activity was strongly inhibited by Mn$^{2+}$, Ni$^{2+}$, and Cu$^{2+}$ ; slightly by Ca$^{2+}$. The purified enzyme produced starch hydrolyzates containing mainly maltose and maltotriose from soluble starch. The starch hydrolyzates were composed of 11% glucose, 59% maltose, 25% maltotriose and 5% maltotetraose.

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.352-358
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• 2002

An amylase that hydrolyzes starch into maltopentaose as a main product was found in the culture supernatant of a strain of Bacillus megaterium KSM B-404 isolated from local soil. The enzyme was purified 129-fold by ammonium sulfate precipitation, DEAE-Toyopearl and Superdex 75 HR 10/30 column using a FPLC system. The molecular weight of the amylase was determined as about 68 kDa by using SDS-PAGE. Optimum pH and temperature of amylase were found to be $50^{\circ}C$ and pH 6.0~7.0, respectively. The enzyme was stable up to $60^{\circ}C$ by addition of $Ca^{2+}$ and its pH stability was in the range of 6.0~10.0. The activity of enzyme was inhibited by $Cu^{2+}$ $Hg^{2+}$ , and $Fe^{3+}$ and maintained by $Ca^{2+}$ and $Mg^{2+}$ . EDTA and pCMB also showed inhibitory effect to the enzyme. TLC and HPLC analysis of the products of the enzyme reaction showed the presence of maltopentaose(52%), maltotriose (25%), maltose (11%), glucose, and maltotetraose in the starch hydrolysates.