A novel amylomaltase from Corynebacterium glutamicum and analysis of the large-ring cyclodextrin products

Srisimarat, Wiraya; Powviriyakul, Areeya; Kaulpiboon, Jarunee; Krusong, Kuakarun; Zimmermann, Wolfgang; Pongsawasdi, Piamsook

doi:10.1007/s10847-010-9890-5

Cited by 40 publications

(91 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The gene encoding the amylomaltase of the mesophilic organism C. glutamicum has been previously characterized (12,14). Recently, CgAM was successfully expressed in E. coli as the recombinant enzyme with His-tag residues at the N terminus and was purified to homogeneity (36). However, after the removal of the His-tag residues by enterokinase, the mature amylomaltase enzyme showed no activity.…”

Section: Discussionmentioning

confidence: 99%

“…Starch transglycosylation activity was measured by the iodine method using soluble potato starch and maltose as the substrates as previously described (36). One unit is defined as the amount of enzyme required to degrade 1 mg starch/ml per min under the described conditions (modified from those described in reference 30).…”

Section: Methodsmentioning

confidence: 99%

“…HPAEC-PAD was used to analyze the LR-CD products with a model ICS 3000 system (Dionex) using a Carbopac PA-100 column (4 by 250 mm; Dionex). A 25-l sample was loaded onto the column and eluted with six continuous-step linear gradients of 200 mM sodium nitrate in 150 mM NaOH at a flow rate of 1 ml/min as previously described (21,36). The size of the LR-CD products was compared with the sizes of the standard LR-CDs that had been confirmed by matrix-assisted laser desorption ionization-time of flight analysis.…”

Section: Methodsmentioning

confidence: 99%

“…In addition, the LR-CD production profile of CgAM was different from that of the wellcharacterized TaAM enzyme (40). By amino acid sequence alignment, the Tyr-172 residue of CgAM was proposed to correspond to the Tyr-54 residue in TaAM (36). The aim of the present study was to investigate the importance of Tyr-172 of CgAM in the LR-CD production profile.…”

mentioning

confidence: 99%

“…The mutation of these residues affected the hydrolysis activity of the TaAM enzyme, which played an important role in product formation (8,9). A novel AM from the mesophilic bacterium Corynebacterium glutamicum ATCC 13032 (CgAM) has recently been reported, and interestingly, it has only a 28% amino acid sequence similarity to TaAM (36). In addition, the LR-CD production profile of CgAM was different from that of the wellcharacterized TaAM enzyme (40).…”

mentioning

confidence: 99%

See 4 more Smart Citations

Altered Large-Ring Cyclodextrin Product Profile Due to a Mutation at Tyr-172 in the Amylomaltase of Corynebacterium glutamicum

Srisimarat

Kaulpiboon

Krusong

et al. 2012

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

c Corynebacterium glutamicum amylomaltase (CgAM) catalyzes the formation of large-ring cyclodextrins (LR-CDs) with a degree of polymerization of 19 and higher. The cloned CgAM gene was ligated into the pET-17b vector and used to transform Escherichia coli BL21(DE3). Site-directed mutagenesis of Tyr-172 in CgAM to alanine (Y172A) was performed to determine its role in the control of LR-CD production. Both the recombinant wild-type (WT) and Y172A enzymes were purified to apparent homogeneity and characterized. The Y172A enzyme exhibited lower disproportionation, cyclization, and hydrolysis activities than the WT. The k cat /K m of the disproportionation reaction of the Y172A enzyme was 2.8-fold lower than that of the WT enzyme. The LR-CD product profile from enzyme catalysis depended on the incubation time and the enzyme concentration. Interestingly, the Y172A enzyme showed a product pattern different from that of the WT CgAM at a long incubation time. The principal LR-CD products of the Y172A mutated enzyme were a cycloamylose mixture with a degree of polymerization of 28 or 29 (CD28 or CD29), while the principal LR-CD product of the WT enzyme was CD25 at 0.05 U of amylomaltase. These results suggest that Tyr-172 plays an important role in determining the LR-CD product profile of this novel CgAM.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Altered Large-Ring Cyclodextrin Product Profile Due to a Mutation at Tyr-172 in the Amylomaltase of Corynebacterium glutamicum

Srisimarat

Kaulpiboon

Krusong

et al. 2012

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

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

Zheng

Zhang

et al. 2019

Starch Stärke

View full text Add to dashboard Cite

The enzyme 4‐α‐glucanotransferase catalyzes the intramolecular and intermolecular transglycosylation of α‐1,4‐glucan, which plays an important role in starch metabolism and starch modification. In this study, the gene encoding a 4‐α‐glucanotransferase from Corallococcus sp. EGB (ccGtase) is cloned and expressed in Escherichia coli BL21 (DE3). CcGtase contains all the conserved amino acid residues of glycoside hydrolase family 77 (GH77), which form the active site environment of the enzymes in the α‐amylase family, while sequence alignments show that 4‐α‐glucanotransferases from myxobacteria constitute an independent group along with the GH77 glucanotransferases. Product analysis shows that CcGtase is able to convert malto‐oligosaccharides and starch into linear maltooligosaccharides with a low degree of polymerization (DP < 12) instead of cycloamylose. The transfer action toward starch is only observed in the presence of glucose, and delayed production of maltose from the transfer reaction is also observed. During the reaction, no apparent hydrolytic activity is observed. Based on the low sequence identity (9–35%) and unique characteristics, CcGtase can be regarded as a new member of the GH77 family.

show abstract

Applications of Starch Debranching Enzymes in Starch Processing

2023

Industrial Starch Debranching Enzymes

View full text Add to dashboard Cite

A novel amylomaltase from Corynebacterium glutamicum and analysis of the large-ring cyclodextrin products

Cited by 40 publications

References 18 publications

Altered Large-Ring Cyclodextrin Product Profile Due to a Mutation at Tyr-172 in the Amylomaltase of Corynebacterium glutamicum

Altered Large-Ring Cyclodextrin Product Profile Due to a Mutation at Tyr-172 in the Amylomaltase of Corynebacterium glutamicum

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

Applications of Starch Debranching Enzymes in Starch Processing

Contact Info

Product

Resources

About