Recently, microbial prodigiosin (PG) has received much attention due to its numerous beneficial applications. The aim of this study was to establish the bioprocessing of marine chitinous wastes (MCWs) for the cost-effective preparation of PG. Of the MCWs, demineralized shrimp shell powders (de-SSP) were found to be a potential source of carbon/nitrogen (C/N) for PG production by bacterial fermentation using Serratia marcescens strains. Further, PG scale-up production was investigated in a 15 L bioreactor system, and the highest yield (6200 mg/L) was achieved during fermentation using 5 L of a novel-designed culture broth that included 1.60% C/N sources (a de-SSP/casein ratio of 7/3), 0.02% K2SO4, and 0.05% K2HPO4, with an initial pH of 6–7. Fermentation was conducted in the dark at 27.5 °C for 8.0 h. This study was the first to report on the utilization of shrimp wastes for cost-effective, large-scale (5 L/pilot) PG production with high productivity (6200 mg/L) in a short cultivation time. The combination of 0.02% K2SO4 and 0.05% K2HPO4 was also found to be a novel salt composition that significantly enhanced PG yield. The red compound was purified and confirmed as PG after analyzing its HPLC profile, mass, and UV/vis spectra. The purified PG was then tested for its bioactivities and showed effective anticancer activities, moderated antioxidant activities, and novel anti-NO effects.
The purpose of this study was to reuse cassava wastewater (CW) for scaled-up production, via the fermentation of prodigiosin (PG), and to conduct an evaluation of its bioactivities. PG was produced at the yield of high 6150 mg/L in a 14 L-bioreactor system, when the designed novel medium (7 L), containing CW and supplemented with 0.25% casein, 0.05% MgSO4, and 0.1% K2HPO4, was fermented with Serratia marcescens TNU01 at 28 °C in 8 h. The PG produced and purified in this study was assayed for some medical effects and showed moderate antioxidant, high anti-NO (anti-nitric oxide), and potential α-glucosidase inhibitory activities. Notably, PG was first reported as a novel effective α-glucosidase inhibitor with a low IC50 value of 0.0183 µg/mL. The commercial anti-diabetic drug acarbose was tested for comparison and had a lesser effect with a high IC50 value of 328.4 µg/mL, respectively. In a docking study, the cation form of PG (cation-PG) was found to bind to the enzyme α-glucosidase by interacting with two prominent amino acids, ASP568 and PHE601, at the binding site on the target enzyme, creating six linkages and showing a better binding energy score (−14.6 kcal/mol) than acarbose (−10.5 kcal/mol). The results of this work suggest that cassava wastewater can serve as a low-cost raw material for the effective production of PG, a potential antidiabetic drug candidate.
α-Amylase inhibitors (aAIs) have been applied for the efficient management of type 2 diabetes. The aim of this study was to search for potential aAIs produced by microbial fermentation. Among various bacterial strains, Pseudomonas aeruginosa TUN03 was found to be a potential aAI-producing strain, and shrimp heads powder (SHP) was screened as the most suitable C/N source for fermentation. P. aeruginosa TUN03 exhibited the highest aAIs productivity (3100 U/mL) in the medium containing 1.5% SHP with an initial pH of 7–7.5, and fermentation was performed at 27.5 °C for two days. Further, aAI compounds were investigated for scaled-up production in a 14 L-bioreactor system. The results revealed a high yield (4200 U/mL) in a much shorter fermentation time (12 h) compared to fermentation in flasks. Bioactivity-guided purification resulted in the isolation of one major target compound, identified as hemi-pyocyanin (HPC) via gas chromatography-mass spectrometry and nuclear magnetic resonance. Its purity was analyzed by high-performance liquid chromatography. HPC demonstrated potent α-amylase inhibitory activity comparable to that of acarbose, a commercial antidiabetic drug. Notably, HPC was determined as a new aAI. The docking study indicated that HPC inhibits α-amylase by binding to amino acid Arg421 at the biding site on enzyme α-amylase with good binding energy (−9.3 kcal/mol) and creating two linkages of H-acceptors.
This study aimed to reuse groundnut oil processing by-product, groundnut cake (GNC) for the low-cost production of prodigiosin (PG) via microbial technology and to assess its novel potential application for the management of black pepper nematodes. Serratia marcescens TUN02 was found as the most active PG-producing strain. Various small-scale experiments conducted in flasks indicated that GNC at 1% may be used as the sole carbon/nitrogen source for cost-effective PG production by fermentation. Notably, no further commercial ingredients and salts are required to supplement into the culture medium of this fermentation. PG was further investigated for scale-up production in a 14-L bioreactor system and PG was produced at high yield (6886 mg/L) with large-scale volume (4 L) in a short cultivation time (10 h). PG was then purified and its nematicidal activity was evaluated and showed effective inhibition of juveniles and egg hatching of Meloidogyne incognita species, harmful on black pepper, with low IC50 values of 0.2 and 0.32 mg/mL, respectively. The simple medium containing 1% GNC is the first report of cost-effective biosynthesis of PG, as well as potential in vitro anti-egg hatching activity of PG. These results indicated the potential application of GNC for low-cost bioproduction of PG for promising and novel use in the management of black pepper nematodes.
A new tirucallane-type triterpenoid igniarine (1), and four known compounds meshimakobnol A (2), meshimakobnol B (3), ergosterol (4) and ergosterol peroxide (5), were purified from the methanol extracts of the fruiting bodies of Phellinus igniarius (DC. ex Fr.) Quél. The structure of 1 was elucidated using a combination of 1D and 2D NMR techniques and HR-ESI-MS analyses. In addition, the isolated compounds were examined for their cytotoxicity against several tumour cell lines and part of the tested compounds demonstrated weak cytotoxicity.
Robusta coffee is a major commercial crop in the Central Highland of Vietnam with high economic and export value. However, this crop is adversely affected by various pathogens, particularly nematodes. This study aimed to screen active anti-nematode rhizobacterial strains for sustainable coffee production. Among more than 200 isolates, the isolate TUN03 demonstrated efficient biocontrol with nearly 100% mortality of J2 coffee nematodes Meloidogyne spp. and 84% inhibition of nematode egg hatching. This active strain was identified as Pseudomonas aeruginosa TUN03 based on its 16S rRNA gene sequence and phylogenetic analysis. In greenhouse tests, the strain TUN03 significantly reduced the coffee nematode population in the rhizome-soil with an 83.23% inhibition rate and showed plant growth-promoting effects. Notably, this is the first report of the nematicidal effect of P. aeruginosa against coffee nematodes. This potent strain further showed an antifungal effect against various crop-pathogenic fungi and was found to be the most effective against Fusarium solani F04 (isolated from coffee roots) with a 70.51% inhibition rate. In addition, high-performance liquid chromatography analysis revealed that this bacterial strain also secretes plant growth regulators including indole acetic acid (IAA), gibberellic acid (GA3), kinetin, and zeatin in significant amounts of 100, 2700, 37, and 9.5 µg/mL, respectively. The data from this study suggest that P. aeruginosa TUN03 may be a potential biocontrol agent and biofertilizer for the sustainable production of Robusta coffee and other crops.
Fishery by-products (FBPs) have been increasingly investigated for the extraction and production of a vast array of active molecules. The aim of this study was to produce phenazine compounds from FBPs via microbial fermentation and assess their novel antinematode effect. Among various FBPs, squid pen powder (SPP) was discovered as the most suitable substrate for phenazine production by Pseudomonas aeruginosa TUN03 fermentation. Various small-scale experiments conducted in flasks for phenazine production indicated that the most suitable was the newly designed liquid medium which included 1% SPP, 0.05% MgSO4, and 0.1% Ca3(PO4)2 (initial pH 7). Phenazines were further studied for scale-up bioproduction in a 14 L bioreactor system resulting in a high yield (22.73 µg/mL) in a much shorter cultivation time (12 h). In the fermented culture broth, hemi-pyocyanin (HPC) was detected as a major phenazine compound with an area percentage of 11.28% in the crude sample. In the bioactivity tests, crude phenazines and HPC demonstrate novel potential nematicidal activity against black pepper nematodes, inhibiting both juveniles (J2) nematodes and egg hatching. The results of this work suggest a novel use of SPP for cost-effective bioproduction of HPC, a novel potential nematodes inhibitor. Moreover, the combination of MgSO4 and Ca3(PO4)2 was also found to be a novel salt composition that significantly enhanced phenazine yield by P. aeruginosa fermentation in this work.
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