Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate
In order to explore the application potential of aspartic protease (Asp) of Trichoderma sp., the protease gene (asp) was cloned from Trichoderma asperellum by real-time polymerase chain reaction, and was successfully expressed in Pichia Pastoris GS115. The recombinant protease (rAsp) was isolated an...
| Main Author: | |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
China Food Publishing Company
2023-10-01
|
| Series: | Shipin Kexue |
| Subjects: | |
| Online Access: | https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-20-021.pdf |
| _version_ | 1797435852445450240 |
|---|---|
| author | ZHOU Di, QIU Xiaoxian, KE Ye, HU Qiuyi |
| author_facet | ZHOU Di, QIU Xiaoxian, KE Ye, HU Qiuyi |
| author_sort | ZHOU Di, QIU Xiaoxian, KE Ye, HU Qiuyi |
| collection | DOAJ |
| description | In order to explore the application potential of aspartic protease (Asp) of Trichoderma sp., the protease gene (asp) was cloned from Trichoderma asperellum by real-time polymerase chain reaction, and was successfully expressed in Pichia Pastoris GS115. The recombinant protease (rAsp) was isolated and purified, and its biochemical properties and its effectiveness in hydrolyzing soy protein isolate (SPI) were studied. The results showed that the protease encoded by the asp gene belonged to the aspartic protease family, and its sequence identity with other members of this family was up to 47.74%. The protease activity of rAsp in the fermentation broth obtained by induced expression in a conical flask was 25.8 U/mL. The optimal reaction pH and temperature of rAsp were 2.5 and 45 ℃, respectively, and rAsp had strong stability in the pH range of 2.0–6.0 and below 45 ℃. The activity of rAsp was promoted by Cu2+ and Mn2+ but inhibited by Fe3+, sodium dodecylsulfate (SDS) and pepstantin. The hydrolysis efficiency of SPI with rAsp was 7.7% higher than that with commercial pepsin. Moreover, the ability of rAsp to reduce the allergenicity of β-conglycinin and glycinin was 1.4 and 1.8 times greater than that of the pepsin, respectively. Therefore, rAsp has potential application in soy protein processing. |
| first_indexed | 2024-03-09T10:54:15Z |
| format | Article |
| id | doaj.art-5b1b7b0cdd0544e9a958297927713289 |
| institution | Directory Open Access Journal |
| issn | 1002-6630 |
| language | English |
| last_indexed | 2024-03-09T10:54:15Z |
| publishDate | 2023-10-01 |
| publisher | China Food Publishing Company |
| record_format | Article |
| series | Shipin Kexue |
| spelling | doaj.art-5b1b7b0cdd0544e9a9582979277132892023-12-01T03:19:24ZengChina Food Publishing CompanyShipin Kexue1002-66302023-10-01442017518210.7506/spkx1002-6630-20230212-105Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein IsolateZHOU Di, QIU Xiaoxian, KE Ye, HU Qiuyi0(Henry Fok School of Biology and Agriculture, Shaoguan University, Shaoguan 512005, China)In order to explore the application potential of aspartic protease (Asp) of Trichoderma sp., the protease gene (asp) was cloned from Trichoderma asperellum by real-time polymerase chain reaction, and was successfully expressed in Pichia Pastoris GS115. The recombinant protease (rAsp) was isolated and purified, and its biochemical properties and its effectiveness in hydrolyzing soy protein isolate (SPI) were studied. The results showed that the protease encoded by the asp gene belonged to the aspartic protease family, and its sequence identity with other members of this family was up to 47.74%. The protease activity of rAsp in the fermentation broth obtained by induced expression in a conical flask was 25.8 U/mL. The optimal reaction pH and temperature of rAsp were 2.5 and 45 ℃, respectively, and rAsp had strong stability in the pH range of 2.0–6.0 and below 45 ℃. The activity of rAsp was promoted by Cu2+ and Mn2+ but inhibited by Fe3+, sodium dodecylsulfate (SDS) and pepstantin. The hydrolysis efficiency of SPI with rAsp was 7.7% higher than that with commercial pepsin. Moreover, the ability of rAsp to reduce the allergenicity of β-conglycinin and glycinin was 1.4 and 1.8 times greater than that of the pepsin, respectively. Therefore, rAsp has potential application in soy protein processing.https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-20-021.pdftrichoderma asperellum; recombinant aspartic protease; biochemical properties; soy protein isolate; allergenicity |
| spellingShingle | ZHOU Di, QIU Xiaoxian, KE Ye, HU Qiuyi Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate Shipin Kexue trichoderma asperellum; recombinant aspartic protease; biochemical properties; soy protein isolate; allergenicity |
| title | Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate |
| title_full | Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate |
| title_fullStr | Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate |
| title_full_unstemmed | Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate |
| title_short | Cloning and Expression of an Aspartic Protease Gene from Trichoderma asperellum and Its Application to the Hydrolysis of Soy Protein Isolate |
| title_sort | cloning and expression of an aspartic protease gene from trichoderma asperellum and its application to the hydrolysis of soy protein isolate |
| topic | trichoderma asperellum; recombinant aspartic protease; biochemical properties; soy protein isolate; allergenicity |
| url | https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-20-021.pdf |
| work_keys_str_mv | AT zhoudiqiuxiaoxiankeyehuqiuyi cloningandexpressionofanasparticproteasegenefromtrichodermaasperellumanditsapplicationtothehydrolysisofsoyproteinisolate |