Identification of active site residues of Fenugreek β-amylase: Chemical modification and in silico approach

Srivastava, Garima; Singh, Vinay Kumar; Kayastha, Arvind M.

doi:10.1016/j.plaphy.2014.08.005

Cited by 4 publications

(4 citation statements)

References 36 publications

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“…Maltose had a non-competitive inhibition pattern on the enzyme, whereas α-cyclodextrin had a competitive pattern, which is consistent with Srivastava et al, 2014 [26].…”

Section: Resultssupporting

confidence: 88%

Biochemical and Thermodynamic Characterization of β amylase from Dioscorea alata L.

Fadunsin

Ebuehi²,

Akande³

et al. 2022

AJRB

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β-amylase (E.C.3.2.1.2) is a starch hydrolyzing enzyme fondly used in foods, pharmaceuticals, and brewing industries to convert starch into maltose. The aim of this study was to determine the physicochemical, kinetic, and thermodynamic properties, as well as the potential industrial use of β-amylase from Dioscorea alata L. Studying the enzyme stability with Arrhenius methods, showed that the enzyme was stable at a temperature range of 20–500C, and had good pH stability by retaining over 50 % of its initial activity over a wide range of pH from 4 – 8 and kinetic stability by increasing the half-life of the enzyme. The activation energy (Ea) for catalysis by water yam β-amylase at 250C was 6.45kcal/mol. The activation energy (Ea), half-life, free energy change (ΔG‡), enthalpy change (ΔH‡), and entropy change (ΔS‡) for inactivation at optimum temperature (400C) and pH 5 were 13.92 kcal/mol, 41.25 min, 20.89 kcal/mol, 13.30 kcal/mol and -24.25 cal/mol/K respectively. Km and Vmax values were reduced from 2.25 to 2.13mg/ml and 2.95 to 1.48 µmol/min/ml respectively. The optimum pH shifted from 5 to 6, while the Optimum temperature increased from 40 to 500C after immobilization. Enzyme retained up to 67 % activity after 5 cycles. The enzyme would be of importance in manufacturing companies based on the kinetics and application features reported in this study since it is a cheap and readily available source.

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“…Maltose had a non-competitive inhibition pattern on the enzyme, whereas α-cyclodextrin had a competitive pattern, which is consistent with Srivastava et al, 2014 [26].…”

Section: Resultssupporting

confidence: 88%

Biochemical and Thermodynamic Characterization of β amylase from Dioscorea alata L.

Fadunsin

Ebuehi²,

Akande³

et al. 2022

AJRB

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“…Thus two substrate molecules bind in tandem in the active site pocket at subsites -2 to -1 and +1 to +2, respectively. It is notable that the term "active site" refers to the amino acid residues involved in the catalysis, along with those involved in interaction with the substrate [59].…”

Section: Discussionmentioning

confidence: 99%

“…6) which is common to all other β-amylases of different origins [50]. According to both structural and functional data [52,56,59,60], three amino-acid residues (Cys, Glu and Asp) conceived to be essential for the catalytic process.…”

Section: Discussionmentioning

confidence: 99%

Identification of a new oat β -amylase by functional proteomics

Halima

Khemakhem

Fendri

et al. 2016

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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“…Moreover, it is easier to modify microbial β-amylase in order to adapt it to flexible and diverse application requirements. Rational and irrational molecular modifications have been used to increase the optimal pH and enhance the catalytic activity of β-amylases [ 18 , 21 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

Expression, biochemical and structural characterization of high-specific-activity β-amylase from Bacillus aryabhattai GEL-09 for application in starch hydrolysis

et al. 2021

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Background β-amylase (EC 3.2.1.2) is an exo-enzyme that shows high specificity for cleaving the α-1,4-glucosidic linkage of starch from the non-reducing end, thereby liberating maltose. In this study, we heterologously expressed and characterized a novel β-amylase from Bacillus aryabhattai. Results The amino acid-sequence alignment showed that the enzyme shared the highest sequence identity with β-amylase from Bacillus flexus (80.73%) followed by Bacillus cereus (71.38%). Structural comparison revealed the existence of an additional starch-binding domain (SBD) at the C-terminus of B. aryabhattai β-amylase, which is notably different from plant β-amylases. The recombinant enzyme purified 4.7-fold to homogeneity, with a molecular weight of ~ 57.6 kDa and maximal activity at pH 6.5 and 50 °C. Notably, the enzyme exhibited the highest specific activity (3798.9 U/mg) among reported mesothermal microbial β-amylases and the highest specificity for soluble starch, followed by corn starch. Kinetic analysis showed that the Km and kcat values were 9.9 mg/mL and 116961.1 s− 1, respectively. The optimal reaction conditions to produce maltose from starch resulted in a maximal yield of 87.0%. Moreover, molecular docking suggested that B. aryabhattai β-amylase could efficiently recognize and hydrolyze maltotetraose substrate. Conclusions These results suggested that B. aryabhattai β-amylase could be a potential candidate for use in the industrial production of maltose from starch.

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Identification of active site residues of Fenugreek β-amylase: Chemical modification and in silico approach

Cited by 4 publications

References 36 publications

Biochemical and Thermodynamic Characterization of β amylase from Dioscorea alata L.

Biochemical and Thermodynamic Characterization of β amylase from Dioscorea alata L.

Identification of a new oat β -amylase by functional proteomics

Expression, biochemical and structural characterization of high-specific-activity β-amylase from Bacillus aryabhattai GEL-09 for application in starch hydrolysis

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