Improvement of Amylomaltase from Thermus aquaticus by Random and Saturation Mutageneses

Fujii, Kazutoshi; Minagawa, Hiroko; Terada, Yukimasa; Takaha, Takeshi; Kuriki, Takashi; Shimada, Jiro; Kaneko, Hiroki

doi:10.5458/jag.52.137

Cited by 8 publications

(9 citation statements)

References 21 publications

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“…This suggests that a yield of LR-CDs with a high degree of polymerization would be improved if only the coupling reaction was inhibited. Fujii et al made amylomaltase mutants by introducing random mutations into the gene coding for this enzyme as proof (71,72). In 2002, it was found that glycogen debranching enzyme (GDE, EC 2.4.1.25/EC 3.2.1.33) from Saccharomyces cerevisiae was also able to produce a LR-CD mixture (73).…”

Section: E Preparation Of Lr-cdsmentioning

confidence: 99%

Large-ring Cyclodextrins

Endō¹

2011

TIGG

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Cyclodextrin (CD) is the common name for cyclic α-1,4-glucans; α-, β-and γ-CD, which are composed of 6, 7, 8 D-glucopyranose units, are well-known. However, most scientists have not recognized large-ring cyclodextrins (LRCDs) composed of more than 9 D-glucopyranose units until recently, although the existence of LR-CDs containing 9-13 D-glucopyranose units in macrocycle was reported about a half century ago. The characteristics of LR-CDs have been studied for the last quarter century, and they are still relatively new materials. In this minireview, the unique characteristics of LR-CDs, including molecular structure, physicochemical properties and inclusion complex formation ability, are discussed. Preparation and purification methods are also reviewed.

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Section: E Preparation Of Lr-cdsmentioning

confidence: 99%

Large-ring Cyclodextrins

Endō¹

2011

TIGG

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“…Tyr54 and Tyr101 278 (T. aquaticus amylomaltase numbering [9]), since both were identified 279 as the aromatic positions involved in the secondary glucan-binding 280 site of the enzyme[49,57,58]. While the former was determined to con-281 trol the hydrolytic activity[57,59], the latter was shown to be responsi-282 ble rather for disproportionating activity of the amylomaltase[58,60].283 Interestingly, the residues of the secondary glucan-binding site from 284 T. aquaticus amylomaltase were found to be not strictly conserved 285 among its counterparts [27,61]. A detailed inspection of the alignment 286 of all 416 4-α-glucanotransferases compared in the present study con-287 firmed that neither Tyr54, nor Tyr101 does represent a conserved posi-288 tion in the family GH77 (Fig.…”

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confidence: 99%

In silico analysis of family GH77 with focus on amylomaltases from borreliae and disproportionating enzymes DPE2 from plants and bacteria

Kuchtová¹,

Janeček²

2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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“…Although the thermostability was high enough to be used for the industrial scale, the yield of cycloamylose using Thermus aquaticus (Taq) amylomaltase decreased to 60% at the end point of the reaction since the enzyme significantly exhibited hydrolytic activity. 47,48) In such a case, we have two ways to overcome the problems. One is enhancing the thermostability of potato D enzyme, and the other is erasing the hydrolytic activity of Taq amylomaltase.…”

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confidence: 99%

“…The yield of cycloamylose using the engineered enzyme remained at more than 90% until the end of reaction, while the reducing power was not significantly increased. 47,48) We are going further to improve enzymes based on the concept of the α amylase family as a rational tool for designing and engineering industrially useful enzymes.…”

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confidence: 99%

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The Concept of the .ALPHA.-Amylase Family: A Rational Tool for Interconverting Glucanohydrolases/Glucanotransferases, and Their Specificities

Kuriki¹,

Takata²,

Yanase³

et al. 2006

J. Appl. Glycosci.

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We found a new enzyme, neopullulanase (EC 3.2.1.135), and showed that it catalyzes the hydrolysis of α-1,4-and α-1,6-glucosidic linkages, as well as transglycosylation to form α-1,4-and α-1,6-glucosidic linkages. Based on the series of experimental results using neopullulanase, we pointed out the same catalytic machinery and the common catalytic mechanism of the enzymes that catalyze these four reactions, and thus, proposed and defined the concept of the α-amylase family. Mutational analyses provided the evidence that one active center of neopullulanase participates in all four reactions; the hydrolysis of α-1,4-and α-1,6-glucosidic linkages and transglycosylation to form α-1,4-and α-1,6-glucosidic linkages. Structural analyses provided the conclusive proof that one active center of neopullulanase participates in all four reactions. We have been trying to interconvert glucanohydrolases glucanotransferases, and their specificities and create tailor-made industrially useful enzymes based on the concept of the α-amylase family. Based on the concept, we engineered Thermus amylomaltase to essentially erase hydrolytic activity and created perfect 4-α-glucanotransferase for the industrial production of cycloamylose. The concept of the α-amylase family is demonstrated here again as a rational tool for interconverting glucanohydrolases glucanotransferases, and their specificities.

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Improvement of Amylomaltase from Thermus aquaticus by Random and Saturation Mutageneses

Cited by 8 publications

References 21 publications

Large-ring Cyclodextrins

Large-ring Cyclodextrins

In silico analysis of family GH77 with focus on amylomaltases from borreliae and disproportionating enzymes DPE2 from plants and bacteria

The Concept of the .ALPHA.-Amylase Family: A Rational Tool for Interconverting Glucanohydrolases/Glucanotransferases, and Their Specificities

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