Optimization of Medium Components and Genes Expression Involved in IAA Biosynthesis by Serratia plymuthica UBCF_13
DOI:
https://doi.org/10.48048/tis.2024.7852Keywords:
HPLC, Indole-3-acetic acid, Medium optimization, Plant growth promoting bacteria, Quantitative reverse transcription-PCRAbstract
Serratia plymuthica UBCF_13 produces the maximum level of indole-3-acetic acid (IAA) in yeast mannitol medium. However, the impact of medium ingredients on gene expression and metabolites related to IAA synthesis remains unclear. Therefore, Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) was used to assess the effects of culture medium optimization components on the genes involved in IAA production. High-Performance Liquid Chromatography (HPLC) has been employed to investigate the impact of medium constituents on the metabolites associated with the biosynthesis of IAA. The highest IAA production was found in sucrose and yeast extract as the carbon and nitrogen sources (134.6 µg/mL). It was also discovered that calcium carbonate and magnesium sulfate are significant inorganic salts for UBCF_13 in the production of IAA. Most genes showed higher levels of expression when sucrose was used as the carbon source, including ipdC, nthA, puuC, amiE, oxdA, tyrB and nthB. Furthermore, the expression of tyrB, puuC, DDC and oxdA was upregulated in response to calcium carbonate, while puuC, nthA, nthB and amiE expression levels were elevated in the presence of magnesium sulfate. Indole-3-acetamide (IAM) was identified with HPLC as an intermediate product in several optimized culture media, with the highest IAA concentration (429.79 µg/mL) which was observed in yeast sucrose medium. Thus, sucrose played a pivotal role in IAA biosynthesis, and yeast sucrose medium supplemented with complete inorganic salts emerged as the optimal medium for IAA production by UBCF_13.
HIGHLIGHTS
- This is the 1st study to evaluate the effect of each medium culture component on the expression of IAA synthesis genes in bacteria
- The expression of several IAA synthesis genes (amiE, nthA, oxdA, tyrB, nthB, puuC and ipdC) was upregulated by using sucrose as the carbon source
- Magnesium sulphate and calcium carbonate were shown to have a significant influence on IAA production, which also increased the expression of several IAA synthesis genes in UBCF_13
- The increase in the expression of genes linked to the IAM pathway (nthA, nthB and amiE) and the identification of IAM as the only intermediate in the extract of optimum medium culture suggest that UBCF_13 may use the IAM pathway to synthesize IAA
- It was discovered that sucrose and yeast extract with inorganic salts (dipotassium phosphate, magnesium sulphate, sodium chloride and calcium carbonate) were the optimal medium composition to produce IAA by UBCF_13
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