Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation
Pullulanase (EC 3.2.1.41) belongs to the amylase family and is often used alone or in combination with other amylases in the industrial production of starch-based products. This enzyme is often required in industrial production because of its better stability. We here truncated the pullulanase gene...
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MDPI AG
2023-02-01
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| author | Xudong Wu Baojie Dou Boyan Wang Mingwang Liu Ruxue Shao Jing Lu Mingsheng Lyu Shujun Wang |
| author_facet | Xudong Wu Baojie Dou Boyan Wang Mingwang Liu Ruxue Shao Jing Lu Mingsheng Lyu Shujun Wang |
| author_sort | Xudong Wu |
| collection | DOAJ |
| description | Pullulanase (EC 3.2.1.41) belongs to the amylase family and is often used alone or in combination with other amylases in the industrial production of starch-based products. This enzyme is often required in industrial production because of its better stability. We here truncated the pullulanase gene from the deep-sea hydrothermal anaerobic archaeon <i>Thermococcus siculi</i> HJ21 and obtained Pul-HJΔ782, which is a member of the α-amylase family GH57. The results revealed that the optimum temperature for Pul-HJΔ782 was 100 °C, and its thermostability at 100 °C improved after truncation. Less than 15% of its enzyme activity was lost after 1 h of incubation at 100 °C, and 57% activity remained after 5 h of treatment. Truncation significantly improved the overall pH tolerance range of Pul-HJΔ782, and its stability in the pH range 4–8 was over 80% relative activity from an average of 60%. The sequence and structural model of Pul-HJΔ782 was analyzed, and its instability index was reduced significantly. Furthermore, the hydrolysates of the truncated and wild-type pullulanase were analyzed, and the enzymatic digestion efficiency of the truncated Pul-HJΔ782 was higher. |
| first_indexed | 2024-03-11T06:48:47Z |
| format | Article |
| id | doaj.art-94d388f3755e4424934a329863ab1a0f |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-11T06:48:47Z |
| publishDate | 2023-02-01 |
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| spelling | doaj.art-94d388f3755e4424934a329863ab1a0f2023-11-17T10:09:38ZengMDPI AGCatalysts2073-43442023-02-0113345310.3390/catal13030453Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by TruncationXudong Wu0Baojie Dou1Boyan Wang2Mingwang Liu3Ruxue Shao4Jing Lu5Mingsheng Lyu6Shujun Wang7Jiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaPullulanase (EC 3.2.1.41) belongs to the amylase family and is often used alone or in combination with other amylases in the industrial production of starch-based products. This enzyme is often required in industrial production because of its better stability. We here truncated the pullulanase gene from the deep-sea hydrothermal anaerobic archaeon <i>Thermococcus siculi</i> HJ21 and obtained Pul-HJΔ782, which is a member of the α-amylase family GH57. The results revealed that the optimum temperature for Pul-HJΔ782 was 100 °C, and its thermostability at 100 °C improved after truncation. Less than 15% of its enzyme activity was lost after 1 h of incubation at 100 °C, and 57% activity remained after 5 h of treatment. Truncation significantly improved the overall pH tolerance range of Pul-HJΔ782, and its stability in the pH range 4–8 was over 80% relative activity from an average of 60%. The sequence and structural model of Pul-HJΔ782 was analyzed, and its instability index was reduced significantly. Furthermore, the hydrolysates of the truncated and wild-type pullulanase were analyzed, and the enzymatic digestion efficiency of the truncated Pul-HJΔ782 was higher.https://www.mdpi.com/2073-4344/13/3/453<i>Thermococcus siculi</i> HJ 21hyperthermia pullulanasetruncationstability |
| spellingShingle | Xudong Wu Baojie Dou Boyan Wang Mingwang Liu Ruxue Shao Jing Lu Mingsheng Lyu Shujun Wang Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation Catalysts <i>Thermococcus siculi</i> HJ 21 hyperthermia pullulanase truncation stability |
| title | Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation |
| title_full | Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation |
| title_fullStr | Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation |
| title_full_unstemmed | Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation |
| title_short | Improved Stability and Hydrolysates of Hyperthermophilic GH57 Type II Pullulanase from the Deep-Sea Archaeon <i>Thermococcus siculi</i> HJ21 by Truncation |
| title_sort | improved stability and hydrolysates of hyperthermophilic gh57 type ii pullulanase from the deep sea archaeon i thermococcus siculi i hj21 by truncation |
| topic | <i>Thermococcus siculi</i> HJ 21 hyperthermia pullulanase truncation stability |
| url | https://www.mdpi.com/2073-4344/13/3/453 |
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