Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast
Strain preservation to maintain stable vitality and the recombinant enzyme activity plays a crucial role in industrial fermentation. A <i>Pichia pastoris</i> strain is routinely stored at −80 °C in a glycerol vial and activated on an antibiotic-containing YPD agar plate before being used...
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MDPI AG
2023-01-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/9/2/104 |
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| author | Xiao Liang Ting Gong Jing-Jing Chen Tian-Jiao Chen Jin-Ling Yang Ping Zhu |
| author_facet | Xiao Liang Ting Gong Jing-Jing Chen Tian-Jiao Chen Jin-Ling Yang Ping Zhu |
| author_sort | Xiao Liang |
| collection | DOAJ |
| description | Strain preservation to maintain stable vitality and the recombinant enzyme activity plays a crucial role in industrial fermentation. A <i>Pichia pastoris</i> strain is routinely stored at −80 °C in a glycerol vial and activated on an antibiotic-containing YPD agar plate before being used for fermentation. Alternatively, the activated strain should be preserved in the agar slant at 2~4 °C (low-temperature storage) for a short period before use. To maximize the utilization of the low-temperature storage for fermentation, we evaluated this method by observing the capacity of both the vitality and the recombinant enzyme activity of the strain at different preservation durations. We found that engineered yeast could be preserved by low-temperature storage for at least 30 months without losing its vitality and biomass enzyme activity by the end of fermentation and could be directly used for the seed cultivation of fermentation, which is more time-saving than strain recovery from −80 °C in a glycerol vial. Moreover, the antibiotic added to the agar slant could be omitted if the heterologous gene was integrated into the host chromosome. Our approach may greatly elevate the production efficiency of the strain. |
| first_indexed | 2024-03-11T08:51:12Z |
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| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-11T08:51:12Z |
| publishDate | 2023-01-01 |
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| series | Fermentation |
| spelling | doaj.art-c9f42e17635149e8a4a600309ae17b5b2023-11-16T20:25:30ZengMDPI AGFermentation2311-56372023-01-019210410.3390/fermentation9020104Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered YeastXiao Liang0Ting Gong1Jing-Jing Chen2Tian-Jiao Chen3Jin-Ling Yang4Ping Zhu5State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaStrain preservation to maintain stable vitality and the recombinant enzyme activity plays a crucial role in industrial fermentation. A <i>Pichia pastoris</i> strain is routinely stored at −80 °C in a glycerol vial and activated on an antibiotic-containing YPD agar plate before being used for fermentation. Alternatively, the activated strain should be preserved in the agar slant at 2~4 °C (low-temperature storage) for a short period before use. To maximize the utilization of the low-temperature storage for fermentation, we evaluated this method by observing the capacity of both the vitality and the recombinant enzyme activity of the strain at different preservation durations. We found that engineered yeast could be preserved by low-temperature storage for at least 30 months without losing its vitality and biomass enzyme activity by the end of fermentation and could be directly used for the seed cultivation of fermentation, which is more time-saving than strain recovery from −80 °C in a glycerol vial. Moreover, the antibiotic added to the agar slant could be omitted if the heterologous gene was integrated into the host chromosome. Our approach may greatly elevate the production efficiency of the strain.https://www.mdpi.com/2311-5637/9/2/104strain preservation<i>Pichia pastori</i>industrial fermentationlow temperature storagevitalityproduction efficiency |
| spellingShingle | Xiao Liang Ting Gong Jing-Jing Chen Tian-Jiao Chen Jin-Ling Yang Ping Zhu Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast Fermentation strain preservation <i>Pichia pastori</i> industrial fermentation low temperature storage vitality production efficiency |
| title | Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast |
| title_full | Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast |
| title_fullStr | Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast |
| title_full_unstemmed | Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast |
| title_short | Influence of Long-Term Agar-Slant Preservation at 4 °C on the Recombinant Enzyme Activity of Engineered Yeast |
| title_sort | influence of long term agar slant preservation at 4 °c on the recombinant enzyme activity of engineered yeast |
| topic | strain preservation <i>Pichia pastori</i> industrial fermentation low temperature storage vitality production efficiency |
| url | https://www.mdpi.com/2311-5637/9/2/104 |
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