Characterization and gene cloning of a maltotriose-forming exo-amylase from Kitasatospora sp. MK-1785

Kamon, Masahiro; Sumitani, Jun-ichi; Tani, Shuji; Kawaguchi, Takashi

doi:10.1007/s00253-015-6396-5

Cited by 24 publications

(9 citation statements)

References 40 publications

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“…The action patterns of SdMFA and SdMFA-CD were assayed using the blue method. , Aliquots of the mixture of 0.2 mL of the enzyme (0.2 nmol mL –1 ) and 1.8 mL of 0.25% (w/v) amylose or amylopectin were incubated at 25 °C. Samples were taken every 10 min to stop the reaction by boiling for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Starch-Binding Domain Modulates the Specificity of Maltopentaose Production at Moderate Temperatures

Ding

Zhao

Han

et al. 2022

J. Agric. Food Chem.

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Maltooligosaccharide-forming amylases (MFAs) hydrolyze starch into maltooligosaccharides with a defined degree of polymerization. However, the enzymatic mechanism underlying the product specificity remains partially understood. Here, we show that Saccharophagus degradans MFA (SdMFA) contains a noncatalytic starch-binding domain (SBD), which belongs to the carbohydrate-binding module family 20 and enables modulation of the product specificity. Removal of SBD from SdMFA resulted in a 3.5-fold lower production of the target maltopentaose. Conversely, appending SBD to another MFA from Bacillus megaterium improved the specificity for maltopentaose. SdMFA exhibited a higher level of exo-action and greater product specificity when reacting with amylopectin than with amylose. Our structural analysis and molecular dynamics simulation suggested that SBD could promote the recognition of nonreducing ends of substrates and delivery of the substrate chain to a groove end toward the active site in the catalytic domain. Furthermore, we demonstrate that a moderate temperature could mediate SBD to interact with the substrate with loose affinity, which facilitates the substrate to slide toward the active site. Together, our study reveals the structural and conditional bases for the specificity of MFAs, providing generalizable strategies to engineer MFAs and optimize the biosynthesis of maltooligosaccharides.

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Section: Methodsmentioning

confidence: 99%

Starch-Binding Domain Modulates the Specificity of Maltopentaose Production at Moderate Temperatures

Ding

Zhao

Han

et al. 2022

J. Agric. Food Chem.

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“…In this study, normal maize starch was gelatinized and hydrolyzed by AMTS into different DHs from 2.0 to 28.5% (Table ). Maltotriose was the predominant sugar produced and PM was 83.1% after 0.25 h hydrolysis, which implies that the exo‐action of AMTS is predominant at the initial stage . The PM gradually decreased and the DH increased with an increase in the hydrolysis time, and it was speculated that the endo‐action of AMTS was more liable to occur with the progression of hydrolysis.…”

Section: Resultsmentioning

confidence: 96%

“…Glucan 1,4‐α‐maltotriohydrolase (AMTS; EC 3.2.1.116) is another addition to the specific amylase with the capacity of both endo‐ and exo‐action . AMTS is also promising in preparing maltotriosyl‐glycosides and maltotriose from starch . To the best of our knowledge, this is the first report on using AMTS and debranching enzyme in RS3‐rich products preparation.…”

Section: Introductionmentioning

confidence: 99%

Effects of α‐maltotriohydrolase hydrolysis prior to debranching on the structure and digestibility of normal maize starch

Zhou

Wei

et al. 2016

Starch Stärke

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Gelatinized normal maize starch was hydrolyzed by glucan 1,4 α‐maltotriohydrolase (AMTS) to various degrees, and then debranched by pullulanase and retrograded at 4°C for 48 h to enhance the formation of type III resistant starch (RS3). AMTS hydrolyzes starch through combined exo‐ and endo‐action, and favors the production of maltotriose. After AMTS hydrolysis, the relative crystallinity and RS contents had all increased, and the highest relative crystallinity (59.2%) and RS content (40.8%) were obtained after 2 h of hydrolysis. The short‐length chains (degree of polymerization (DP) 13–30) and long‐length chains (DP > 130) were nearly simultaneously hydrolyzed by AMTS; thus, medium‐length chains with a DP of 30–130 accumulated after moderate hydrolysis. The results suggest that RS3 formation is affected by branch chain lengths, especially medium‐length chains (DP 30–130), and it can be improved when AMTS hydrolysis precedes debranching.

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“…KO-8940 (PseG5A) and maltohexaose-forming amylase from Corallococcus sp. EGB (CorG6A) for comparison (Table 1; Fujita et al, 1989;Shida et al, 1992;Doukyu et al, 2007;Kamon et al, 2015;Li et al, 2015). Four conserved sequence regions (Region I to IV) that had been discovered and used to define α-amylases in glycosyl hydrolase family 13 were found in all 8 amylase targets (Figure 4C; Janeček, 2002).…”

Section: Structure Analysis Of Sdg5amentioning

confidence: 99%

Carbohydrate-Binding Module and Linker Allow Cold Adaptation and Salt Tolerance of Maltopentaose-Forming Amylase From Marine Bacterium Saccharophagus degradans 2-40T

et al. 2021

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Marine extremophiles produce cold-adapted and/or salt-tolerant enzymes to survive in harsh conditions. These enzymes are naturally evolved with unique structural features that confer a high level of flexibility, solubility and substrate-binding ability compared to mesophilic and thermostable homologs. Here, we identified and characterized an amylase, SdG5A, from the marine bacterium Saccharophagus degradans 2-40T. We expressed the protein in Bacillus subtilis and found that the purified SdG5A enabled highly specific production of maltopentaose, an important health-promoting food and nutrition component. Notably, SdG5A exhibited outstanding cold adaptation and salt tolerance, retaining approximately 30 and 70% of its maximum activity at 4°C and in 3 M NaCl, respectively. It converted 68 and 83% of starch into maltooligosaccharides at 4 and 25°C, respectively, within 24 h, with 79% of the yield being the maltopentaose. By analyzing the structure of SdG5A, we found that the C-terminal carbohydrate-binding module (CBM) coupled with an extended linker, displayed a relatively high negative charge density and superior conformational flexibility compared to the whole protein and the catalytic domain. Consistent with our bioinformatics analysis, truncation of the linker-CBM region resulted in a significant loss in activities at low temperature and high salt concentration. This highlights the linker-CBM acting as the critical component for the protein to carry out its activity in biologically unfavorable condition. Together, our study indicated that these unique properties of SdG5A have great potential for both basic research and industrial applications in food, biology, and medical and pharmaceutical fields.

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Characterization and gene cloning of a maltotriose-forming exo-amylase from Kitasatospora sp. MK-1785

Cited by 24 publications

References 40 publications

Starch-Binding Domain Modulates the Specificity of Maltopentaose Production at Moderate Temperatures

Starch-Binding Domain Modulates the Specificity of Maltopentaose Production at Moderate Temperatures

Effects of α‐maltotriohydrolase hydrolysis prior to debranching on the structure and digestibility of normal maize starch

Carbohydrate-Binding Module and Linker Allow Cold Adaptation and Salt Tolerance of Maltopentaose-Forming Amylase From Marine Bacterium Saccharophagus degradans 2-40T

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