Biotechnology and molecular biology of the ?-glucosidase inhibitor acarbose

Wehmeier, Udo F.; Piepersberg, Wolfgang

doi:10.1007/s00253-003-1477-2

Cited by 189 publications

(185 citation statements)

References 55 publications

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“…SE50/110 encodes a mixture of proteins related to enzymes of the primary and secondary metabolism (27), supporting a new type of starch metabolism based on acarbose as a central structure. Acarbose serves as an inhibitor for starch-degrading enzymes used by competitors in the natural environment and also serves as an acceptor molecule for glucose and longer oligosaccharides (16,27). Acarbose is structurally related to starch molecules ( Figure 3), and acarbose metabolism involves several enzymes closely related to starch-modifying enzymes (16).…”

Section: Discussionmentioning

confidence: 99%

“…Acarbose serves as an inhibitor for starch-degrading enzymes used by competitors in the natural environment and also serves as an acceptor molecule for glucose and longer oligosaccharides (16,27). Acarbose is structurally related to starch molecules ( Figure 3), and acarbose metabolism involves several enzymes closely related to starch-modifying enzymes (16). Since our data show that a single amino acid change in the active site of a CGTase is sufficient to make it an acarbose-modifying enzyme (and vice versa), the R-amylase encoded by the acarbose (acb) gene of the Actinoplanes sp.…”

Section: Discussionmentioning

confidence: 99%

“…This secondary metabolite is produced industrially using engineered strains of Actinoplanes sp. SE50/110 (16), and it is used as a drug (Glucobay/Precose) in the treatment of diabetes patients to slow the intestinal release of glucose (17). Acarbose is a strong reversible inhibitor of many R-amylase family enzymes, including the cyclization reactions with starch catalyzed by the CGTases of Bacillus circulans strain 251 [K i ) 0.2 µM (18)] and Thermoanaerobacterium thermosulfurigenes strain EM1 [Tabium; 1 …”

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Single Amino Acid Mutations Interchange the Reaction Specificities of Cyclodextrin Glycosyltransferase and the Acarbose-Modifying Enzyme Acarviosyl Transferase

2004

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Single amino acid mutations interchange the reaction specificities of cyclodextrin glycosyltransferase and the acarbose-modifying enzyme acarviosyl transferase Leemhuis, H.; Wehmeier, U.F.; Dijkhuizen, Lubbert Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. D-42079 Wuppertal, Germany ReceiVed May 15, 2004; ReVised Manuscript ReceiVed July 22, 2004 ABSTRACT: Acarviosyl transferase (ATase) from Actinoplanes sp. SE50/110 is a bacterial enzyme that transfers the acarviosyl moiety of the diabetic drug acarbose to sugar acceptors. The enzyme exhibits 42% sequence identity with cyclodextrin glycosyltransferases (CGTase), and both enzymes are members of the R-amylase family, a large clan of enzymes acting on starch and related compounds. ATase is virtually inactive on starch, however. In contrast, ATase is the only known enzyme to efficiently use acarbose as substrate (2 µmol min -1 mg -1 ); acarbose is a strong inhibitor of CGTase and of most other R-amylase family enzymes. This distinct reaction specificity makes ATase an interesting enzyme to investigate the variation in reaction specificity of R-amylase family enzymes. Here we show that a G140H mutation in ATase, introducing the typical His of the conserved sequence region I of the R-amylase family, changed ATase into an enzyme with 4-R-glucanotransferase activity (3.4 µmol min -1 mg -1 ). Moreover, this mutation introduced cyclodextrin-forming activity into ATase, converting 2% of starch into cyclodextrins. The opposite experiment, removing this typical His side chain in CGTase (H140A), introduced acarviosyl transferase activity in CGTase (0.25 µmol min -1 mg -1 ).The R-amylase family, or glycoside hydrolase family 13 (1), is a large family of enzymes acting on R-glycosidic bonds in starch and related compounds (2). About 20 different reaction specificities have been identified in this family, including hydrolysis of R-(1,4)-and R-(1,6)-glycosidic linkages (e.g., R-amylase and isoamylase, respectively), as well as the formation of R-(1,4)-and R-(1,6)-glycosidic bonds (e.g., amylosucrase, acarviosyl transferase, cyclodextrin glycosyltransferase, and branching enzyme, respectively; Figure 1) (2, 3). All members use an R-retaining double displacement mechanism (4) in which reactions proceed via a covalent glycosyl-enzyme intermediate (5). Glycosidic bond cleavage occurs between subsites -1 and +1 ( Figure 2A), and after cleavage the glycosyl reaction intermediate re...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Single Amino Acid Mutations Interchange the Reaction Specificities of Cyclodextrin Glycosyltransferase and the Acarbose-Modifying Enzyme Acarviosyl Transferase

2004

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“…The 3 mL reaction mixture contained 13 mM 2-oxoglutarate, 1.7 mM NADH or NADPH. 15 mM NH 4 Cl, 15 mM NH 3 •H 2 O, or 7.5 mM (NH 4 ) 2 SO 4 was added separately for the reductive GDH activity. Reactions were started by adding the enzyme.…”

Section: Methodsmentioning

confidence: 99%

“…As a competitive α-glucosidase inhibitor, acarbose is widely used in the therapy of noninsulin-dependent diabetes mellitus owing to its good therapeutic and non-toxic effects [2,3].…”

Section: Introductionmentioning

confidence: 99%

Influence of Ammonium Salt and Fermentation pH on Acarbose Yield from <i>Streptomyces</i> M37

Ren¹,

Chen²,

Tong³

2015

Trop. J. Pharm Res

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From Gene to Product: The Advantage of Integrative Biotechnology

Schmidt

2006

Handbook of Pharmaceutical Biotechnology

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PRODUCTION ORGANISMS AND EXPRESSION SYSTEMSDesign and development of all microbial production processes start with the selection of appropriate organisms, strains, and expression systems enabling high yields and high quality of a desired product with defi ned pharmacological properties. Industrially Established Recombinant Expression SystemsIndustrially established expressions systems for production of the marketed compounds are, besides inclusion, body-forming Escherichia coli strains, the yeast Saccharomyces cerevisiae and mammalian cells like CHO-and BHK-cells (Table 1.1-1). These systems were the genetically and physiologically most advanced and therefore mostly applied when recombinant production processes were starting to be developed in the mid-1980s and are now widely accepted by regulatory bodies. E. coli and S. cerevisiae can be grown cheaply and rapidly, are amenable to high cell density fermentations with biomasses of up to 130 g/L, possess short generation times, have high capacities to accumulate foreign proteins, are easy to handle, and are established fermentation organisms.However, because gene recombinant pharmaceuticals continuously gain an increasing importance in medicine and are expected to help curing diseases that are not yet treatable today, new expression systems have to be exploited enabling the production of such pharmaceuticals with innovative properties that simultaneously meet key criteria like consistent product quality and cost effectiveness. Of particular interest in this regard are expression systems enabling the secretion of TABLE 1.1-1. Industrially Used Recombinant Expression Systems

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Biotechnology and molecular biology of the ?-glucosidase inhibitor acarbose

Cited by 189 publications

References 55 publications

Single Amino Acid Mutations Interchange the Reaction Specificities of Cyclodextrin Glycosyltransferase and the Acarbose-Modifying Enzyme Acarviosyl Transferase

Single Amino Acid Mutations Interchange the Reaction Specificities of Cyclodextrin Glycosyltransferase and the Acarbose-Modifying Enzyme Acarviosyl Transferase

Influence of Ammonium Salt and Fermentation pH on Acarbose Yield from <i>Streptomyces</i> M37

From Gene to Product: The Advantage of Integrative Biotechnology

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