Proteomic analysis of the caffeine response in Cordyceps militaris

Name: Thanyarat Puttika

keyword: Cordyceps militaris

MeSH: Caffeine

Classification :.DDC: e-Theses

; Caffeine

MeSH: Adenosine deaminase -- 307324

MeSH: Proteomics

LCSH: Caffeine

LCSH: Adenosine deaminase -- 307324

LCSH: Proteomics

Abstract: This study is aimed to address the caffeine response of entomopathogenic fungi, Cordyceps militaris, involved in growth and development, metabolite production and caffeine degradation. Our results demonstrated that caffeine showed a dosage effect on the growth, development, and adenosine production of C. militaris. Caffeine concentrations (0, 1 and 10 mM) inhibited the mycelial growth of C. militaris, with a reduction of mycelial growth rate at 1 mM (0.37±0.01 cm/day) and 10mM (0.27±0.01 cm/day) compared to without caffeine (0.42±0.02 cm/day) and the number of conidia was also reduced by 23% and 77%, respectively compared to control. Caffeine also affected the fungal metabolite production by increasing the extracellular adenosine productivity 5-fold (from 4.34±0.60 µg/g/day to 24.28±0.65 µg/g/day) but there was no change in cordycepin (p<0.05). Interestingly, this fungus could metabolize caffeine, resulting in the accumulation of its derivatives, theophylline (55.23±2.18 µg/mL), theobromine (16.95±0.97 µg/mL), and paraxanthine (12.88±1.22µg/mL). These findings provide a new understanding of how caffeine affects the growth of C. militaris. This information could be useful to produce the bioactive compounds either adenosine or caffeine derivatives. Next, intracellular caffeine-regulated C. militaris proteins were analyzed and identified using MALDI-TOF-MS/MS. Then, all Differentially Expressed Proteins (DEPs) were classified into coherent groups. Total 491 proteins discovered to be implicated in the caffeine response, 42 of these proteins showed upregulation, whereas 449 showed downregulation. With 1 mM caffeine, 334 DEPs were upregulated and 284 DEPs were downregulated. At 10 mM, 285 DEPs were upregulated, and 333 DEPs were downregulated. In low and high caffeine concentrations, the majority of DEPs were associated with ion binding, nucleotide binding, and transferase activity. Here, the log fold change treatment with 1 and 10 mM of caffeine demonstrated a significant difference (p<0.001). Fourteen DEPs were upregulated out of a total of 63 DEPs, including (1) cell wall structure and synthesis (e.g. actin cytoskeleton regulatory protein, beta 1, 3 endoglucanase, and beta 1, 3 glucosyltransferase), (2) transporters (MFS transporter), (3) primary and secondary metabolite production (acyltransferase, acyl hydrolase, and lysophospholipase), (4) molybdopterin cofactor of oxidoreductases and (5) redox enzyme NAD(P)H that may require xanthine dehydrogenase involved in the caffeine degradation pathway as well as in stress response. In contrast, the downregulated proteins were demonstrated to be involved in cell wall synthesis (e.g. chitin synthase export chaperone), cell cycle-related proteins (e.g. Sir2 family protein, and protein kinase domain-containing protein), RNA processing and translation (e.g. ribosomal RNA-processing protein and translation elongation factor), ABC multidrug transporter, cellular respiratory chain (e.g. D-lactate dehydrogenase and citrate synthase).

Mae Fah Luang University. Learning Resources and Educational Media Center

Address: CHIANG RAI

Email: library@mfu.ac.th

Name: Amorn Owatworakit

Role: Advisor

Created: 2023

Modified: 2024-01-17

Issued: 2024-01-17

วิทยานิพนธ์/Thesis

application/pdf

CallNumber: e-Theses

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biological Sciences

Grantor: Mae Fah Luang University

©copyrights Mae Fah Luang University

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