High maltose-forming, Ca2+-independent and acid stable α-amylase from a novel acidophilic bacterium, Bacillus acidicola

Sharma, Archana; Satyanarayana, T.

doi:10.1007/s10529-010-0322-9

Cited by 47 publications

(23 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The commonly used industrial A. oryzae a-amylase produced syrups of 50-60% maltose, whereas the recently published high maltose-forming a-amylase from Bacillus acidicola produced 67% maltose [28]. Under the experimental conditions, 69% maltose was produced by both Anoxybacillus a-amylases, and this percentage is comparable to those of the A. oryzae and Bacillus acidicola amylases.…”

Section: Analysis Of Hydrolysis Productsmentioning

confidence: 72%

See 1 more Smart Citation

Cloning and characterization of two new thermostable and alkalitolerant α-amylases from the Anoxybacillus species that produce high levels of maltose

Chai

Rahman

Illias

et al. 2012

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

Two genes that encode α-amylases from two Anoxybacillus species were cloned and expressed in Escherichia coli. The genes are 1,518 bp long and encode 506 amino acids. Both sequences are 98% similar but are distinct from other well-known α-amylases. Both of the recombinant enzymes, ASKA and ADTA, were purified using an α-CD-Sepharose column. They exhibited an optimum activity at 60°C and pH 8. Both amylases were stable at pH 6-10. At 60°C in the absence of Ca²⁺, negligible reduction in activity for up to 48 h was observed. The activity half-life at 65°C was 48 and 3 h for ASKA and ADTA, respectively. In the presence of Ca²⁺ ions, both amylases were highly stable for at least 48 h and had less than a 10% decrease in activity at 70°C. Both enzymes exhibited similar end-product profiles, and the predominant yield was maltose (69%) from starch hydrolysis. To the best of our knowledge, most α-amylases that produce high levels of maltose are active at an acidic to neutral pH. This is the first report of two thermostable, alkalitolerant recombinant α-amylases from Anoxybacillus that produce high levels of maltose and have an atypical protein sequence compared with known α-amylases.

show abstract

Section: Analysis Of Hydrolysis Productsmentioning

confidence: 72%

“…a-Amylases that produce high levels of maltose are usually active at acidic to neutral pH values [19,28]. In this study, the two Anoxybacillus a-amylases are thermostable and alkalitolerant and produce high levels of maltose.…”

Section: Introductionmentioning

confidence: 86%

Cloning and characterization of two new thermostable and alkalitolerant α-amylases from the Anoxybacillus species that produce high levels of maltose

Chai

Rahman

Illias

et al. 2012

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

show abstract

“…Bacillus acidicola was isolated from a soil sample collected from Rameshwaram (Tamil Nadu, India) (Sharma and Satyanarayana 2010) and routinely grown on starch/ peptone/beef extract/agar (g l -1 :10 g starch, 5 g peptone, 3 g beef extract, 20 g agar and pH 4.5) at 37°C and preserved at -20°C in glycerol. The culture is deposited at Microbial Type Culture Collection, Institute of Microbial Technology, Chandigarh, India (MTCC-10504).…”

Section: Organism and Culture Conditionsmentioning

confidence: 99%

“…Determination of a-amylase activity and protein concentration a-Amylase was assayed according to Sharma and Satyanarayana (2010). One unit of saccharogenic a-amylase activity is defined as the amount of enzyme that liberates 1 lmol reducing sugar as maltose min -1 under the assay conditions.…”

Section: Gene Cloning and Plasmid Constructionmentioning

confidence: 99%

See 1 more Smart Citation

Cloning and expression of acidstable, high maltose-forming, Ca2+-independent α-amylase from an acidophile Bacillus acidicola and its applicability in starch hydrolysis

Sharma

Satyanarayana

2012

Extremophiles

Self Cite

View full text Add to dashboard Cite

The α-amylase encoding gene from acidophilic bacterium Bacillus acidicola was cloned into pET28a(+) vector and expressed in Escherichia coli BL21 (DE3). The recombinant E. coli produced a 15-fold higher α-amylase than B. acidicola strain. The recombinant α-amylase was purified to homogeneity by one-step nickel affinity chromatography using Ni(2+)-NTA resin with molecular mass of 62 KDa. It is active in the pH range between 3.0 and 7.0 and 30 and 100 °C with optimum at pH 4.0 and 60 °C. The enzyme is Ca(2+)-independent with K (m) and k (cat) values (on soluble starch) of 1.6 mg ml(-1) and 108.7 s(-1), respectively. The α-amylase of B. acidicola is acidstable, high maltose forming and Ca(2+)-independent, and therefore, is a suitable candidate for starch hydrolysis and baking.

show abstract

Engineering of Bacillus amyloliquefaciens α‐Amylase with Improved Calcium Independence and Catalytic Efficiency by Error‐Prone PCR

Tian

Dong

et al. 2017

Starch Stärke

View full text Add to dashboard Cite

Bacillus amyloliquefaciens α‐amylase (BAA) is one of the most important amylases and presents a wide range of applications in many processes involving starch‐liquefaction. However, its activity and thermostability largely depend on the existence of calcium ions, which is a major obstacle for its wide‐scale industrial applications. In the present study, to enhance the calcium independence of a mesophilic α‐amylase from B. amyloliquefaciens CICIM B2125, a novel calcium‐independent mutant Q264S is evolved by directed evolution using error‐prone polymerase chain reaction (PCR). Enzymatic properties and kinetic parameters of mutant Q264S were subsequently analyzed. Although the thermostability at 60°C and optimum temperature of Q264S are slightly decreased, its calcium independence and catalytic efficiency are significantly improved, with no apparent changes of specific activity and optimum pH compared with those of the wild‐type enzyme. Furthermore, three‐dimensional structure analysis showed that the improved calcium independence and catalytic efficiency of Q264S are most likely to be a subtle balance among the hydrophobic interactions, electrostatic forces, and hydrogen bonds around calcium‐binding sites and active‐site regions. To the best of our knowledge, this is the first report on increasing calcium independence of BAA by error‐prone PCR. These results could aid the understanding of mechanistic bases of calcium independence of BAA and thus facilitate the design and engineering of more efficient and robust calcium‐independent BAA variants.

show abstract

High maltose-forming, Ca2+-independent and acid stable α-amylase from a novel acidophilic bacterium, Bacillus acidicola

Cited by 47 publications

References 11 publications

Cloning and characterization of two new thermostable and alkalitolerant α-amylases from the Anoxybacillus species that produce high levels of maltose

Cloning and characterization of two new thermostable and alkalitolerant α-amylases from the Anoxybacillus species that produce high levels of maltose

Cloning and expression of acidstable, high maltose-forming, Ca2+-independent α-amylase from an acidophile Bacillus acidicola and its applicability in starch hydrolysis

Engineering of Bacillus amyloliquefaciens α‐Amylase with Improved Calcium Independence and Catalytic Efficiency by Error‐Prone PCR

Contact Info

Product

Resources

About