Gene Expression and Biochemical Characterization of a GH77 4‐α‐Glucanotransferase CcGtase From <i>Corallococcus</i> sp. EGB

Li, Zhoukun; Zheng, Wenjie; Zhang, Lei; Wang, Yanxin; Zhang, Yajuan; Qiao, Yan; Luo, Xue; Huang, Yan; Cui, Zhongli

doi:10.1002/star.201800254

Cited by 5 publications

(4 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Among them, are the maltohexaose-forming α-amylase AmyM from Corallococcus sp. EGB [ 8 ] and the possible G1 + G2-producing α-amylase Amy1 from Massilia timonae CTI-57 [ 28 ], which show sequences identities of 42% and 51% with AmyCf, respectively. Otherwise, AmyCf has low sequence identity (<35%) with other 16 members of subfamily GH13_6.…”

Section: Resultsmentioning

confidence: 99%

“…Then, we transformed P. pastoris GS115 with the pEFαA-amyCf-based plasmid and isolated the positive transformants which could grow on YPD plates containing 100 mg/L zeocin (Invitrogen, R25001). Inducible expression of the amylase AmyCf in P. pastoris was performed as described before [ 8 ]. Briefly, isolated transformants were grown in 50 mL YPD medium (1% yeast extract, 2% peptone and 2% glucose) at 30 °C and 200 rpm for 24 h, followed by transferring cells into 100 mL of BMMY medium (1% yeast extract, 2% peptone, 1% glycerol, 1.34% YNB, 0.1 mol/L phosphate buffer and 0.04 mg/L biotin).…”

Section: Methodsmentioning

confidence: 99%

“…EGB [ 3 ], maltotriose-producing amylase (G3) from Kitasatospora [ 4 ], maltotetraose-producing amylase (G4) from Bacillus halodurans MS-2–5 [ 5 ], maltopentaose-forming amylase (G5) from Bacillus stearothermophilus [ 6 ], maltohexaose-forming amylase [ 7 ] and GH77 4-α-glucanotransferase CcGtase from Corallococcus sp. EGB [ 8 ]. To achieve efficient enzymatic hydrolyzation, pre-treatment of raw starch under higher temperature is needed to destroy the internal ordered structure of starch particles.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification and Characterization of Novel Malto-Oligosaccharide-Forming Amylase AmyCf from Cystobacter sp. Strain CF23

Wang,

Zhang,

Wang

et al. 2023

Foods

View full text Add to dashboard Cite

Malto-oligosaccharides (MOSs) from starch conversion is advantageous for food and pharmaceutical applications. In this study, an efficient malto-oligosaccharide-forming α-amylase AmyCf was identified from myxobacter Cystobacter sp. strain CF23. AmyCf is composed of 417 amino acids with N-terminal 41 amino acids as the signal peptide, and conserved glycoside hydrolase family 13 (GH13) catalytic module and predicted C-terminal domain with β-sheet structure are also identified. Phylogenetic and functional analysis demonstrated that AmyCf is a novel member of GH13_6 subfamily. The special activity of AmyCf toward soluble starch and raw wheat starch is 9249 U/mg and 11 U/mg, respectively. AmyCf has broad substrate specificity toward different types of starches without requiring Ca2+. Under ideal circumstances of 60 °C and pH 7.0, AmyCf hydrolyzes gelatinized starch into maltose and maltotriose and maltotetraose as the main hydrolytic products with more than 80% purity, while maltose and maltotriose are mainly produced from the hydrolysis of raw wheat starch with more than 95% purity. The potential applicability of AmyCf in starch processing is highlighted by its capacity to convert gelatinized starch and raw starch granules into MOSs. This enzymatic conversion technique shows promise for the low-temperature enzymatic conversion of raw starch.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Identification and Characterization of Novel Malto-Oligosaccharide-Forming Amylase AmyCf from Cystobacter sp. Strain CF23

Wang,

Zhang,

Wang

et al. 2023

Foods

View full text Add to dashboard Cite

show abstract

“…The amount of reducing sugar was determined spectrophotometrically at 540 nm. A unit of enzyme activity was defined as the amount of enzyme that was required to release reducing sugars equivalent to 1 μmol of maltose per min under the test conditions 15 . We determined the protein concentration by the Bradford method, and bovine serum albumin was used as the standard 17 …”

Section: Methodsmentioning

confidence: 99%

Phenylalanine 314 is essential for the activity of maltogenic amylase from Corallococcus sp. EGB

Zhong

Xia

et al. 2021

Biotech and App Biochem

View full text Add to dashboard Cite

Maltogenic amylase CoMA from Corallococcus sp. strain EGB catalyzes the hydrolysis and transglycosylation of maltooligosaccharides and soluble starch into maltose, the sole hydrolysate. This process yields pure maltose with potentially wide applications. Here, we identified and evaluated the role of phenylalanine 314 (F314), a key amino acid located near the active center, in the catalytic activities of the CoMA. Site‐directed mutagenesis analysis showed that the activity of a F314L mutant on potato starch substrate decreased to 26% of that of wild‐type protein. Compared with the wild‐type, F314L exhibited similar substrate specificity, hydrolysis pattern, pH, and temperature requirements. Circular dichroism spectrum data showed that the F314L mutation did not affect the structure of the folded protein. In addition, kinetic analysis demonstrated that F314L exhibited an increased Km value with lower substrate affinity. Homology modeling showed that the benzene ring structure of F314L was involved in π–π conjugation, which might potentially affect the affinity of CoMA toward starch. Taken together, these data demonstrated that F314 is essential for the hydrolytic activity of the CoMA from Corallococcus sp. strain EGB.

show abstract

Novel Malto‐Oligosaccharide‐Producing Amylase AmyAc from Archangium sp. Strain AC19 and Its Catalytic Properties

et al. 2021

View full text Add to dashboard Cite

Functional maltooligosaccharides (MOSs) derived from starch have potential applications in the food industry and medicine due to their specific functional properties. In this study, a novel maltose (G2) and maltotriose (G3)-producing amylase AmyAc from Archangium sp. strain AC19 is identified. AmyAc is a protein of 874 amino acids, with an N-terminal signal peptide as well as hydrolase family 13 (GH13) catalytic and C-terminal binding modules. However, its sequence is not highly similar to any of the reported starch hydrolases. The V max of AmyAc is 124 µmol min −1 mg −1 under optimal conditions of 50°C and pH 7.0. AmyAc catalyzes the conversion of MOSs and soluble starch into maltose and maltotriose as the end-product with more than 85% purity. AmyAc initially produces G2, G3, and oligosaccharides with even units, following by the release of G2 and oligosaccharides with odd units during later stages of hydrolysis, indicating a distinct catalytic selectivity. The production of MOSs from starch by AmyAc is of significant interest due to potential applications in starch processing.

show abstract

Gene Expression and Biochemical Characterization of a GH77 4‐α‐Glucanotransferase CcGtase From Corallococcus sp. EGB

Cited by 5 publications

References 40 publications

Identification and Characterization of Novel Malto-Oligosaccharide-Forming Amylase AmyCf from Cystobacter sp. Strain CF23

Identification and Characterization of Novel Malto-Oligosaccharide-Forming Amylase AmyCf from Cystobacter sp. Strain CF23

Phenylalanine 314 is essential for the activity of maltogenic amylase from Corallococcus sp. EGB

Novel Malto‐Oligosaccharide‐Producing Amylase AmyAc from Archangium sp. Strain AC19 and Its Catalytic Properties

Contact Info

Product

Resources

About