Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95.
Alpha-amylase is a very important enzyme in the starch conversion process. Most of the α-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extr...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4374950?pdf=render |
| _version_ | 1818850980881498112 |
|---|---|
| author | Liang Xian Fei Wang Xiang Luo Yu-Liang Feng Jia-Xun Feng |
| author_facet | Liang Xian Fei Wang Xiang Luo Yu-Liang Feng Jia-Xun Feng |
| author_sort | Liang Xian |
| collection | DOAJ |
| description | Alpha-amylase is a very important enzyme in the starch conversion process. Most of the α-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extracellular amylolytic enzyme was purified from the culture broth of newly isolated Talaromyces pinophilus strain 1-95. The purified amylolytic enzyme, with an apparent molecular weight of 58 kDa on SDS-PAGE, hydrolyzed maltopentaose, maltohexaose, and maltoheptaose into mainly maltose and maltotriose and minor amount of glucose, confirming the endo-acting mode of the enzyme, and hence, was named Talaromyces pinophilus α-amylase (TpAA). TpAA was most active at pH 4.0-5.0 (with the temperature held at 37°C) and 55°C (at pH 5.0), and stable within the pH range of 5.0-9.5 (at 4°C) and below 45°C (at pH 5.0). Interestingly, the Ca2+ did not improve its enzymatic activity, optimal temperature, or thermostability of the enzyme, indicating that the TpAA was Ca2+-independent. TpAA displayed higher enzyme activity toward malto-oligosaccharides and dextrin than other previously reported α-amylases. This highly active Ca2+-independent α-amylase may have potential applications in starch-to-ethanol conversion process. |
| first_indexed | 2024-12-19T06:57:45Z |
| format | Article |
| id | doaj.art-903baac153b84219ab84c0a70ed2a5fd |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T06:57:45Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-903baac153b84219ab84c0a70ed2a5fd2022-12-21T20:31:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01103e012153110.1371/journal.pone.0121531Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95.Liang XianFei WangXiang LuoYu-Liang FengJia-Xun FengAlpha-amylase is a very important enzyme in the starch conversion process. Most of the α-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extracellular amylolytic enzyme was purified from the culture broth of newly isolated Talaromyces pinophilus strain 1-95. The purified amylolytic enzyme, with an apparent molecular weight of 58 kDa on SDS-PAGE, hydrolyzed maltopentaose, maltohexaose, and maltoheptaose into mainly maltose and maltotriose and minor amount of glucose, confirming the endo-acting mode of the enzyme, and hence, was named Talaromyces pinophilus α-amylase (TpAA). TpAA was most active at pH 4.0-5.0 (with the temperature held at 37°C) and 55°C (at pH 5.0), and stable within the pH range of 5.0-9.5 (at 4°C) and below 45°C (at pH 5.0). Interestingly, the Ca2+ did not improve its enzymatic activity, optimal temperature, or thermostability of the enzyme, indicating that the TpAA was Ca2+-independent. TpAA displayed higher enzyme activity toward malto-oligosaccharides and dextrin than other previously reported α-amylases. This highly active Ca2+-independent α-amylase may have potential applications in starch-to-ethanol conversion process.http://europepmc.org/articles/PMC4374950?pdf=render |
| spellingShingle | Liang Xian Fei Wang Xiang Luo Yu-Liang Feng Jia-Xun Feng Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95. PLoS ONE |
| title | Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95. |
| title_full | Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95. |
| title_fullStr | Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95. |
| title_full_unstemmed | Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95. |
| title_short | Purification and characterization of a highly efficient calcium-independent α-amylase from Talaromyces pinophilus 1-95. |
| title_sort | purification and characterization of a highly efficient calcium independent α amylase from talaromyces pinophilus 1 95 |
| url | http://europepmc.org/articles/PMC4374950?pdf=render |
| work_keys_str_mv | AT liangxian purificationandcharacterizationofahighlyefficientcalciumindependentaamylasefromtalaromycespinophilus195 AT feiwang purificationandcharacterizationofahighlyefficientcalciumindependentaamylasefromtalaromycespinophilus195 AT xiangluo purificationandcharacterizationofahighlyefficientcalciumindependentaamylasefromtalaromycespinophilus195 AT yuliangfeng purificationandcharacterizationofahighlyefficientcalciumindependentaamylasefromtalaromycespinophilus195 AT jiaxunfeng purificationandcharacterizationofahighlyefficientcalciumindependentaamylasefromtalaromycespinophilus195 |