Transcriptome Analysis Reveals the Variations in Enzyme Production of Saccharopolyspora rosea A22 under Different Temperatures
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Materials
2.2. Cultivation and Morphological Observation of Saccharopolyspora rosea A22
2.3. Measurement of Biomass
2.4. Production of Cooked Wheat Qu
- (1)
- Wheat pre-treatment: High-quality red skin wheat was selected to make wheat Qu and the wheat grains were kept dry. The wheat was screened at the conveyor port of the wheat mill to remove impurities and ensure cleanliness. It was then crushed using a wheat mill, with the roller gap set at 1.7–1.8 mm, resulting in 2 to 3 pieces of wheat per grain to break the wheat peel and expose the starch in the wheat grain.
- (2)
- Moistening material cooking: Approximately 40% of normal temperature pure water (by total mass) was added to moisten the material. Care was taken to add water and wheat in batches to avoid local areas becoming too wet or too dry. The mixture was then placed into a pre-heated boiling steam box and steamed for about 40 min.
- (3)
- Cooling inoculation: The digester was opened, and the material was discharged to reduce the temperature to 45 °C and remove any caking material. All cultures were mixed well and an appropriate volume of cultures (5% of the total mass) was weighed using a clean measuring cylinder and evenly added to the material.
- (4)
- Wheat Qu culture: The mixed wheat Qu was placed in an incubator and the fermentation temperature was set to 37 °C, 42 °C and 50 °C, respectively. The culture was maintained for 7 d, and after fermentation was complete, the wheat Qu was fully dried for use.
2.5. Determination of Enzyme Activity in Cooked Wheat Qu
2.6. Transcriptome Sequencing and Analysis
2.7. Primer Design and Gene Expression Verification
2.8. Statistical Analysis
3. Results and Discussion
3.1. Morphological Comparison of Saccharopolyspora rosea A22 at Different Temperatures
3.2. Biomass Differences in Saccharopolyspora rosea A22 at Different Temperatures
3.3. Comparison of Enzyme Activity in Preparation of Cooked Wheat Qu by Saccharopolyspora rosea A22 at Different Temperatures
3.4. Transcriptome Sequencing Gene Statistics and Sample Relationship
3.5. Gene Expression Analysis
3.6. Functional Annotation of Differentially Expressed Genes
3.6.1. GO Analysis
3.6.2. KEGG Analysis
3.7. Verification of Key Genes
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene ID | Name | Sequence (5′ to 3′) |
---|---|---|
Amy1F(NV263_RS18035) | Amy1F | CGCTGACCTGAAAACCGAA |
Amy1R | TGACCGTTGAAGGTGTAGC | |
Amy2F(NV263_RS12515) | Amy2F | ATGGCTTCCTCGCTGGTCA |
Amy2R | GTTGCTGCCCTCCTTGTCC | |
ClpXF(NV263_RS05520) | ClpXF | GATGAGCCCGTACTTGATC |
ClpXR | CGGCTGCGGCAAGACCTAC | |
DnaB(NV263_RS30285) | DnaBF | GCTACCACGGCTCCGAGGG |
DnaBR | CGGGAGAACTCGGAGACTT |
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Lin, C.; Lu, P.; Ma, J.; Li, Z.; Han, X.; Ji, Z.; Liu, S.; Mao, J. Transcriptome Analysis Reveals the Variations in Enzyme Production of Saccharopolyspora rosea A22 under Different Temperatures. Foods 2024, 13, 2696. https://doi.org/10.3390/foods13172696
Lin C, Lu P, Ma J, Li Z, Han X, Ji Z, Liu S, Mao J. Transcriptome Analysis Reveals the Variations in Enzyme Production of Saccharopolyspora rosea A22 under Different Temperatures. Foods. 2024; 13(17):2696. https://doi.org/10.3390/foods13172696
Chicago/Turabian StyleLin, Congyu, Peiqi Lu, Jingqiu Ma, Zhihui Li, Xiao Han, Zhongwei Ji, Shuangping Liu, and Jian Mao. 2024. "Transcriptome Analysis Reveals the Variations in Enzyme Production of Saccharopolyspora rosea A22 under Different Temperatures" Foods 13, no. 17: 2696. https://doi.org/10.3390/foods13172696