Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris
A Pichia pastoris expression method for the efficient extracellular production of Procambarus clarkii invertebrate-type lysozyme 1 (pc-iLys1) was established. First, the synthetic codon-optimized cDNA encoding pc-iLys1 was expressed in the P. pastoris SMD1168 with a molecular mass of about 17.1 kDa....
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Zhejiang University Press
2019-10-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.10.081 |
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| author | SHUI Yan GUAN Zhengbing YE Junxian SHI Yonghong LIU Guofeng XU Zenghong |
| author_facet | SHUI Yan GUAN Zhengbing YE Junxian SHI Yonghong LIU Guofeng XU Zenghong |
| author_sort | SHUI Yan |
| collection | DOAJ |
| description | A Pichia pastoris expression method for the efficient extracellular production of Procambarus clarkii invertebrate-type lysozyme 1 (pc-iLys1) was established. First, the synthetic codon-optimized cDNA encoding pc-iLys1 was expressed in the P. pastoris SMD1168 with a molecular mass of about 17.1 kDa. The expression condition in a shaking flask was optimized and the best condition for pc-iLys1 production was as follow: the cells were cultured in pH 7.0 and grown at 28℃ for 96 h supplemented with 1.5% methanol. High cell density fermentation was then performed using a fed-batch strategy in a 5 L fermenter. Extracellular lytic activity and dry cell mass concentration reached to 2 052.6 U/mL and 98.1 g/L after 120 h, respectively. The purified recombinant pc-iLys1 exhibited extensive antimicrobial activities toward the selected bacteria (including Gram-positive and Gram-negative bacteria). In a word, the P. pastoris expression strain and the method established in this work are promising for the industrial high-yield production of pc-iLys1 protein, and the recombinant pc-iLys1 has potential as an antimicrobial agent for treating the infectious diseases of P. clarkii. |
| format | Article |
| id | doaj-art-8ee20b50d36642e7a1f8dafdf663c7f3 |
| institution | DOAJ |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2019-10-01 |
| publisher | Zhejiang University Press |
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| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-8ee20b50d36642e7a1f8dafdf663c7f32025-08-20T03:16:14ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552019-10-014552653210.3785/j.issn.1008-9209.2018.10.08110089209Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastorisSHUI YanGUAN ZhengbingYE JunxianSHI YonghongLIU GuofengXU ZenghongA Pichia pastoris expression method for the efficient extracellular production of Procambarus clarkii invertebrate-type lysozyme 1 (pc-iLys1) was established. First, the synthetic codon-optimized cDNA encoding pc-iLys1 was expressed in the P. pastoris SMD1168 with a molecular mass of about 17.1 kDa. The expression condition in a shaking flask was optimized and the best condition for pc-iLys1 production was as follow: the cells were cultured in pH 7.0 and grown at 28℃ for 96 h supplemented with 1.5% methanol. High cell density fermentation was then performed using a fed-batch strategy in a 5 L fermenter. Extracellular lytic activity and dry cell mass concentration reached to 2 052.6 U/mL and 98.1 g/L after 120 h, respectively. The purified recombinant pc-iLys1 exhibited extensive antimicrobial activities toward the selected bacteria (including Gram-positive and Gram-negative bacteria). In a word, the P. pastoris expression strain and the method established in this work are promising for the industrial high-yield production of pc-iLys1 protein, and the recombinant pc-iLys1 has potential as an antimicrobial agent for treating the infectious diseases of P. clarkii.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.10.081invertebrate-type lysozyme<italic>Procambarus clarkii</italic><italic>Pichia pastoris</italic>expression condition optimizationantimicrobial activity |
| spellingShingle | SHUI Yan GUAN Zhengbing YE Junxian SHI Yonghong LIU Guofeng XU Zenghong Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris 浙江大学学报. 农业与生命科学版 invertebrate-type lysozyme <italic>Procambarus clarkii</italic> <italic>Pichia pastoris</italic> expression condition optimization antimicrobial activity |
| title | Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris |
| title_full | Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris |
| title_fullStr | Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris |
| title_full_unstemmed | Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris |
| title_short | Efficient extracellular expression and antimicrobial activity of Procambarus clarkii invertebrate-type lysozyme in Pichia pastoris |
| title_sort | efficient extracellular expression and antimicrobial activity of procambarus clarkii invertebrate type lysozyme in pichia pastoris |
| topic | invertebrate-type lysozyme <italic>Procambarus clarkii</italic> <italic>Pichia pastoris</italic> expression condition optimization antimicrobial activity |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.10.081 |
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