Microbes as a tool for the bioremediation of fish waste from the environment and the production of value-added compounds: a review.

Fish are the most edible protein source worldwide and generate several remnants such as scales, viscera, head, bone, and skin. Fish wastes are not disposed of properly, which adversely affects the environment, especially the water bodies where fish processing industries dispose of their waste. Fish waste mainly contains nitrogen, oil, fat, salts, heavy metals, and organic compounds, which increase the biological oxygen demand and chemical oxygen demand. Fish waste can degrade in various ways, such as physicochemical or by enzymatic action, but using microbes is an environmentally friendly approach that can provide valuable compounds such as products such as collagen, chitin, minerals, and fish protein concentrates. This review is designed to focus on the suitability of microbes as tools for fish waste degradation and the production of certain associated. This study also provides insight into the production of other compounds such as protease, chitinase, and chitin applicability of these products. After processing, fish waste as a microbial growth media for enzyme production since microorganisms synthesize enzymes such as proteases, protein hydrolysates, lipids, and chitinase, which have broader applications in the pharmaceutical, cosmetic, biomedical material, and food processing industries.

Enhanced vanillin production from eugenol by Bacillus cereus NCIM-5727.

Biovanillin production by a wild strain of Bacillus cereus NCIM-5727 is studied using eugenol as the precursor aiming to achieve maximum vanillin productivity. Based on shake flask optimization, molar yield and global volumetric productivity of vanillin reached up to 71.2% (6.6 gL-1) and 0.18 g(Lh)-1, respectively, at 36 h by resting cells of B. cereus NCIM-5727 at the optimum cell concentration of 3 gL-1 using eugenol concentration of 10 gL-1 at 37 ºC, buffer pH 7.0, buffer volume 10%, and shaking speed 180 rpm. Furthermore, small-scale optimization in a bioreactor at the controlled aeration rate of 0.5 Lmin-1, agitation rate of 210 rpm, and pH 7.0 enhanced the global volumetric productivity of vanillin up to 0.28 g(Lh)-1 at 25 h of bioconversion. The highest vanillin molar yield (75.2%) is reported using resting cells of B. cereus NCIM-5727 upon eugenol biotransformation and found stable for 10 h.

A rapid and simple ultra high performance liquid chromatography method for the simultaneous determination of methoxyphenol derivatives involved in the eugenol catabolic pathway.

An efficient ultra high performance liquid chromatography method of separation was developed for the analysis of six important methoxyphenol derivatives involved in the eugenol catabolic pathway. In the present study, an Acquity UPLC BEH C18 column was used for the chromatographic separation of the industrially important phenolic compounds such as vanillin, vanillic acid, ferulic acid, coniferyl alcohol, and coniferyl aldehyde obtained during microbial transformation of eugenol. Eluted components were identified using the dual wavelength (254 and 310 nm) UV detector. A gradient method of elution using mobile phase of aqueous 1 mM trifluoroacetic acid (Solvent A) and methanol (Solvent B) at a flow rate of 0.3 mL/min separated all the five intermediate methoxyphenol derivatives along with their precursor eugenol within 15 min with stable baseline resolution. Method validation was performed for the accurate quantification of vanillin, coniferyl aldehyde, and eugenol using the parameters of linearity, specificity, precision, limit of detection, limit of quantification, and robustness. The developed method would be helpful for clear separation and identification of the five most important intermediate metabolites of the eugenol catabolism pathway.

Decolorization and degradation of reactive orange 16 by Bacillus stratosphericus SCA1007.

Efficient bacterial strain was isolated from the dye contaminated area and identified as Bacillus stratosphericus SCA1007 based on 16S rRNA gene sequence (GenBank under accession number KY992944). This isolate was selected based on its potential to efficiently decolorize reactive orange 16 dye which is extensively used in textile industries. Various culture conditions like dye concentration, temperature, pH, salinity, and additional nitrogen source were optimized in the present study. The optimal conditions for decolorization of reactive orange 16 was found to be: dye concentration 150 mg/L, pH 7, temperature 35 °C, and yeast extract as nitrogen source. The isolate was also resistant to 4% saline culture condition. Decolorization and degradation of dye were confirmed through UV-visible spectroscopy, Fourier transform infrared (FTIR) and liquid chromatography-mass spectrometry analysis (LC-MS). Toxicity studies were performed on Escherichia coli and Vigna radiata to confirm the non-toxic nature of the degraded metabolites. This is the first study demonstrating complete decolorization of reactive orange 16 dye by Bacillus stratosphericus SCA1007 at high salinity within 10 h of incubation under optimized conditions.

Mutational analysis of microbial hydroxycinnamoyl-CoA hydratase-lyase (HCHL) towards enhancement of binding affinity: A computational approach.

Improving the industrial enzyme for better yield of the product is important and a challenging task. One of such important industrial enzymes is microbial Hydroxycinnamoyl-CoA hydratase-lyase (HCHL). It converts feruloyl-CoA to vanillin. We place our efforts towards the improvement of its catalytic activity with comprehensive computational investigation. Catalytic core of the HCHL was explored with molecular modeling and docking approaches. Site-directed mutations were introduced in the catalytic site of HCHL in a sequential manner to generate different mutants of HCHL. Basis of mutation is to increase the interaction between HCHL and substrate feruloyl-CoA through interatomic forces and hydrogen bond formation. A rigorous molecular dynamics (MD) simulation was performed to check the stability of mutant's structure. Root mean square deviation (RMSD), root mean square fluctuation (RMSF), dynamic cross correlation (DCCM) and principal component analysis (PCA) were also performed to analyze flexibility and stability of structures. Docking studies were carried out between different mutants of HCHL and feruloyl-CoA. Investigation of the different binding sites and the interactions with mutant HCHLs and substrate allowed us to highlight the improved performance of mutants than wild type HCHL. This was further validated with MD simulation of complex consisting of different mutants and substrate. It further confirms all the structures are stable. However, mutant-2 showed better affinity towards substrate by forming hydrogen bond between active site and feruloyl-CoA. We propose that increase in hydrogen bond formation might facilitate in dissociation of vanillin from feruloyl-CoA. The current work may be useful for the future development of 'tailor-made' enzymes for better yield of vanillin.