Next Article in Journal
Danger Control Programs Cause Tissue Injury and Remodeling
Previous Article in Journal
Current Status and Future Perspectives of Mass Spectrometry Imaging
Int. J. Mol. Sci. 2013, 14(6), 11302-11318; doi:10.3390/ijms140611302
Article

A High Molecular-Mass Anoxybacillus sp. SK3-4 Amylopullulanase: Characterization and Its Relationship in Carbohydrate Utilization

1
, 2
, 1
, 1
 and 1,*
Received: 9 April 2013; in revised form: 3 May 2013 / Accepted: 14 May 2013 / Published: 28 May 2013
(This article belongs to the Section Bioactives and Nutraceuticals)
View Full-Text   |   Download PDF [1001 KB, uploaded 19 June 2014]   |   Browse Figures
Abstract: An amylopullulanase of the thermophilic Anoxybacillus sp. SK3-4 (ApuASK) was purified to homogeneity and characterized. Though amylopullulanases larger than 200 kDa are rare, the molecular mass of purified ApuASK appears to be approximately 225 kDa, on both SDS-PAGE analyses and native-PAGE analyses. ApuASK was stable between pH 6.0 and pH 8.0 and exhibited optimal activity at pH 7.5. The optimal temperature for ApuASK enzyme activity was 60 °C, and it retained 54% of its total activity for 240 min at 65 °C. ApuASK reacts with pullulan, starch, glycogen, and dextrin, yielding glucose, maltose, and maltotriose. Interestingly, most of the previously described amylopullulanases are unable to produce glucose and maltose from these substrates. Thus, ApuASK is a novel, high molecular-mass amylopullulanase able to produce glucose, maltose, and maltotriose from pullulan and starch. Based on whole genome sequencing data, ApuASK appeared to be the largest protein present in Anoxybacillus sp. SK3-4. The α-amylase catalytic domain present in all of the amylase superfamily members is present in ApuASK, located between the cyclodextrin (CD)-pullulan-degrading N-terminus and the α-amylase catalytic C-terminus (amyC) domains. In addition, the existence of a S-layer homology (SLH) domain indicates that ApuASK might function as a cell-anchoring enzyme and be important for carbohydrate utilization in a streaming hot spring.
Keywords: Anoxybacillus; amylase; Bacillus; Geobacillus; glycoside hydrolase 13; pullulan; pullulanase; starch; thermostable enzyme Anoxybacillus; amylase; Bacillus; Geobacillus; glycoside hydrolase 13; pullulan; pullulanase; starch; thermostable enzyme
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Export to BibTeX |
EndNote


MDPI and ACS Style

Kahar, U.M.; Chan, K.-G.; Salleh, M.M.; Hii, S.M.; Goh, K.M. A High Molecular-Mass Anoxybacillus sp. SK3-4 Amylopullulanase: Characterization and Its Relationship in Carbohydrate Utilization. Int. J. Mol. Sci. 2013, 14, 11302-11318.

AMA Style

Kahar UM, Chan K-G, Salleh MM, Hii SM, Goh KM. A High Molecular-Mass Anoxybacillus sp. SK3-4 Amylopullulanase: Characterization and Its Relationship in Carbohydrate Utilization. International Journal of Molecular Sciences. 2013; 14(6):11302-11318.

Chicago/Turabian Style

Kahar, Ummirul M.; Chan, Kok-Gan; Salleh, Madihah M.; Hii, Siew M.; Goh, Kian M. 2013. "A High Molecular-Mass Anoxybacillus sp. SK3-4 Amylopullulanase: Characterization and Its Relationship in Carbohydrate Utilization." Int. J. Mol. Sci. 14, no. 6: 11302-11318.


Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert