Purification and characterization of a novel α-amylase from a newly isolated Bacillus methylotrophicus strain P11-2

Xie, Fuhong; Quan, Shujing; Liu, Dehai; Ma, Huan; Li, Feng; Fuzhong, Zhou; Chen, Guocan

doi:10.1016/j.procbio.2013.09.025

Cited by 67 publications

(54 citation statements)

References 43 publications

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“…). These results clearly showed that the enzyme cleaved internal α‐1,4 linkages and a similar pattern of soluble starch hydrolysis has been reported by Xie et al . The end products of soluble starch hydrolysis by the enzyme indicated the amylase from Anoxybacillus sp.…”

Section: Resultssupporting

confidence: 88%

Purification and characterization of a novel and versatile α-amylase from thermophilicAnoxybacillussp. YIM 342

et al. 2015

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An extracellular thermophilic a-amylase (1, 4-a-D-glucanohydrolase, EC 3.2.1.l) from Anoxybacillus sp. YIM 342 was partially purified by ultrafiltration followed by ammonium sulfate fractionation and dialysis. This procedure was followed by a single purification step using gel filtration chromatography to give a 10.41% yield, 1912.2 U/mg specific activity, and 32-fold purification enrichment. The molecular mass of the purified a-amylase was determined to be 68 kDa using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimum temperature and pH of the a-amylase activity were 80°C and 9.0, respectively. Furthermore, the high hydrolysis rate toward amylose and amylopectin suggested the enzyme has significant potential for applications in starch degradation. The K m and V max of the amylase toward soluble starch was 4.18 mg/mL and 7.48 mmol/min/mg, respectively. This enzyme hydrolyzes soluble starch to glucose, maltose, and maltotriose, indicating that the amylase represents a promising candidate for applications in the biofuel industry.

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Section: Resultssupporting

confidence: 88%

Purification and characterization of a novel and versatile α-amylase from thermophilicAnoxybacillussp. YIM 342

et al. 2015

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“…On the other hand, GpA1 was activated by Ca 2+ and Triton X‐100. This result is in agreement with previous reports that most α‐amylases require calcium for activity or stability .…”

Section: Resultssupporting

confidence: 93%

A novel saccharifying α‐amylase of Antarctic psychrotolerant fungi Geomyces pannorum: Gene cloning, functional expression, and characterization

et al. 2015

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A DNA segment encoding a novel a-amylase was cloned from the Antarctic psychrotolerant fungi Geomyces pannorum. The gene has an open reading frame of 1761 bp, deduced 586 amino acids, revealing a relatively low homology to a-amylases from diverse fungal genera. The a-amylase gene was overexpressed in Pichia pastoris, and the recombinant a-amylase was purified and characterized. Enzymatic activity was optimal at pH 6.0 and 70°C, and the specific activity was 9.78 Â 10 3 U/mg when using soluble starch as substrate. Moreover, the enzyme retained more than 90% of its maximum activity over a wide pH range (6.0-9.0). Exceptionally, the main hydrolysis products of soluble starch catalyzed by the enzyme were glucose and maltose, of which glucose reached a concentration of 68.8% (w/w). This is the first report on identifying thermophilic fungal amylase from psychrotolerant fungi. Also, the preliminary study on the high-glucose-producing a-amylase with detailed enzymatic properties shows a potential application prospect in syrup industry.

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“…Similar to Bacillus methylotrophicus strain P11‐2‐Mg +2 acts as cofactor and Hg +2 completely inhibited amylase activity. Fe +3 also inhibited .…”

Section: Resultsmentioning

confidence: 90%

“…). The amylase from B. methylotrophicus P11‐2, also had an optimum of pH 7 and showed much the same response to lower and higher pH . Fincan et.…”

Section: Resultsmentioning

confidence: 93%

Production optimization, purification and characterization of α-amylase from thermophilicBacillus licheniformis TSI-14

Shukla

Singh

2015

Starch - Stärke

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Bacillus licheniformis TSI‐14 was isolated form a natural hot water reservoir of TulsiShyam in the Gir Forest, Gujarat (India). The bacterium was identified by 16S rDNA gene sequencing. Amylase production was optimized using an RSM approach (Response Surface Methodology; variables: temperature, pH, medium yeast extract and peptone, and incubation period). These data were refined using Central Composite Design (CCD). The conditions deduced for the maximal α‐amylase production were 50°C, pH 7.0, and incubation times of 2 days. A 4.6 fold increase in α‐amylase was achieved. The molecular weight of the purified enzyme is 31 KDa by SDS‐PAGE. It is active at elevated temperatures such as 90°C, and it prefers pH values around neutral. Optimum catalysis and stability occurred at 70°C and pH 7. There is a strong Mg++ dependency evident; other metal ions show inhibition to a lesser or greater extent. The chelator EDTA completely inhibits while inhibitors urea and beta‐mercaptoethanol show inhibition to a greater extent; thiourea enhances activity. Malto–oligosaccharides are intermediate products along with maltose as expected of a α‐amylase.

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Purification and characterization of a novel α-amylase from a newly isolated Bacillus methylotrophicus strain P11-2

Cited by 67 publications

References 43 publications

Purification and characterization of a novel and versatile α-amylase from thermophilicAnoxybacillussp. YIM 342

Purification and characterization of a novel and versatile α-amylase from thermophilicAnoxybacillussp. YIM 342

A novel saccharifying α‐amylase of Antarctic psychrotolerant fungi Geomyces pannorum: Gene cloning, functional expression, and characterization

Production optimization, purification and characterization of α-amylase from thermophilicBacillus licheniformis TSI-14

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