ABSTRACT
Marine biofouling causes huge economic losses to the marine industry every year. Albofungin is a potential antifoulant showing strong anti-macrofouling activities against larval settlement of major fouling organisms. In the present study, directed RNA-seq and proteomic analyses were used to investigate changes in the transcriptome and proteome of a major fouling barnacle Amphibalanus amphitrite cyprids in response to albofungin treatment. Results showed that albofungin treatment remarkably upregulated the metabolism of xenobiotics by the cytochrome P450 pathway to discharge the compound and downregulated energy metabolic processes. Intriguingly, immunostaining and whole-mount in situ hybridization (WISH) revealed the spatial expression patterns of selected differentially expressed genes (glutathione S-transferase [GST], nitric oxide synthase [NOS], and calmodulin [CaM]) distributed in the thorax and antennule of A. amphitrite. Our study provides new insights into the mechanism of albofungin in interrupting the larval settlement of A. amphitrite and suggests its potential application as an antifouling agent in marine environments.
ABSTRACT
The genus Streptomyces is known to harbor numerous biosynthetic gene clusters (BGCs) of potential utility in synthetic biology applications. However, it is often difficult to link uncharacterized BGCs with the secondary metabolites they produce. Proteomining refers to the strategy of identifying active BGCs by correlating changes in protein expression with the production of secondary metabolites of interest. In this study, we devised a shotgun proteomics-based workflow to identify active BGCs during fermentation when a variety of compounds are being produced. Mycelia harvested during the non-producing growth phase served as the background. Proteins that were differentially expressed were clustered based on the proximity of the genes in the genome to highlight active BGCs systematically from label-free quantitative proteomics data. Our software tool is easy-to-use and requires only 1 point of comparison where natural product biosynthesis was significantly different. We tested our proteomining clustering method on three Streptomyces species producing different compounds. In Streptomyces coelicolor A3(2), we detected the BGCs of calcium-dependent antibiotic, actinorhodin, undecylprodigiosin, and coelimycin P1. In Streptomyces chrestomyceticus BCC24770, 7 BGCs were identified. Among them, we independently re-discovered the type II PKS for albofungin production previously identified by genome mining and tedious heterologous expression experiments. In Streptomyces tenebrarius, 5 BGCs were detected, including the known apramycin and tobramycin BGC as well as a newly discovered caerulomycin A BGC in this species. The production of caerulomycin A was confirmed by LC-MS and the inactivation of the caerulomycin A BGC surprisingly had a significant impact on the secondary metabolite regulation of S. tenebrarius. In conclusion, we developed an unbiased, high throughput proteomics-based method to complement genome mining methods for the identification of biosynthetic pathways in Streptomyces sp.
ABSTRACT
Depletion of fossil fuels and environmental problems are encouraging research on alternative fuels of renewable sources. Biodiesel is a promising alternative fuel to be used as a substitute to the petroleum based diesel fuels. However, the cost of biodiesel production is high and is attributed mainly to the feedstock used which leads to the investigation of low cost feedstocks that are economically feasible. In this paper, we report on the utilization of lipid obtained from food waste as a low-cost feedstock for biodiesel production. Lipid from food waste was transesterified with methanol using base and lipase as catalysts. The maximum biodiesel yield was 100% for the base (KOH) catalyzed transesterification at 1:10M ratio of lipid to methanol in 2h at 60°C. Novozyme-435 yielded a 90% FAME conversion at 40°C and 1:5 lipid to methanol molar ratio in 24h. Lipid obtained from fungal hydrolysis of food waste is found to be a suitable feedstock for biodiesel production.