Dechlorination of wastewater from shell-based glucosamine processing by mangrove wetland-derived fungi.

Wastewater from processing crustacean shell features ultrahigh chloride content. Bioremediation of the wastewater is challenging due to the high chloride ion content, making it inhospitable for most microorganisms to survive and growth. In this study, mangrove wetland-derived fungi were first tested for their salt tolerance, and the highly tolerant isolates were cultured in shrimp processing wastewater and the chloride concentration was monitored. Notably, the filamentous fungal species Aspergillus piperis could remove over 70% of the chloride in the wastewater within 3 days, with the fastest biomass increase (2.01 times heavier) and chloride removal occurring between day one and two. The chloride ions were sequestered into the fungal cells. The genome of this fungal species contained Cl- conversion enzymes, which may have contributed to the ion removal. The fungal strain was found to be of low virulence in larval models and could serve as a starting point for further considerations in bioremediation of shell processing wastewater, promoting the development of green technology in the shell processing industry.

[Biochemical properties of Scedosporium aurantiacum extracellular elastase-like protease].

Extracellular elastase-like protease is one of the key virulence proteases of Scedosporium aurantiacum. To date, little is known about this enzyme in terms of genetic information, structure, properties and virulence mechanism due to the difficulties in purification caused by its low secretion amount, high specific activity, uncompleted genome sequencing and annotation. This work investigated the gene, structure and enzymatic properties of this enzyme. The S. aurantiacum elastase-like protease from the fungal culture supernatant was analyzed through tandem mass spectrometry (MS/MS) approach, illustrating its primary structure. Bioinformatics tools were employed to predict the conserved domain and tertiary structure, the enzymatic properties were also studied. It turned out that S. aurantiacum extracellular elastase-like protease demonstrated well hydrolysis towards elastin and bovine achilles tendon collagen, with Vmax of 18.14 µg/s and 17.57 µg/s respectively, better than fish scale gelatin, with the lowest hydrolysis effect on casein. Its activity towards elastin was lower than that of the elastase from porcine pancreas, with values of Kcat/Km of 3.541 (µg/s) and 4.091 (µg/s), respectively. It was an alkaline protease, with optimal pH 8.2 and temperature 37 oC. Zn2+ promoted the enzymatic activity while Ca2+, Mg2+, Na+, elastatinal and PMSF inhibited its activity. Its sequence was similar to Paecilomyces lilacinus secreted serine protease (PDB Entry: c3f7oB_) with multiple conserved fractions each containing more than 7 amino acids, thus suitable for design of PCR primer. This study increased our knowledge on S. aurantiacum extracellular elastase-like protease in terms of structure and enzymatic properties, and may facilitate later studies on protein expression and virulence mechanism.

Composition and content analysis of fluoride in inorganic salts of the integument of Antarctic krill (Euphausia superba).

Ash of Antarctic krill integument (AAKI) was prepared by sintering the integument at 550°C under air atmosphere for 4 hours, and its composition was analyzed by X-ray diffraction (XRD), fourier transform infrared spectroscopy (FTIR) and electron dispersive spectroscopy (EDS). XRD results showed that the major phase in AAKI was ascribed to apatite. Besides, it was noticed that the (300) peak of AAKI shifted to 33.07°, which was coincident with that of fluorapatite (FA). The FTIR results confirmed the presence of phosphate ions, and the absence of -OH. The EDS results confirmed the presence of Ca, P, O and F elements in the ash sample. The content of FA in the ash was determined to be 50.4%, and the proportion of fluorine in the form of FA to the total fluorine in the integument was 40.5%. Based on the XRD, FTIR and EDS results, it can be concluded that FA was the main form of fluoride in the integument of Antarctic krill.

Localization of trypsin-like protease in postmortem tissue of white shrimp (Litopenaeus vannamei) and its effect in muscle softening.

Fluorescein-isothiocyanate (FITC) labeled trypsin-like protease was prepared and injected into the hepatopancreas of white shrimp. Different segments of the injected shrimp were analyzed with a fluorescence microscope during storage. FITC-trypsin-like protease can be detected in the first segment of shrimp muscle at day 4, while it cannot be observed in the second segment until day 6. The results showed that trypsin-like protease can migrate from hepatopancreas to the tail portion. Texture profile analysis showed that soybean trypsin inhibitor retarded the softening of the shrimp muscle. The rheological results revealed that the content of myosin heavy chain (MHC) in shrimp muscle was decreased with the extended storage time. Proteomics analysis displayed that trypsin-like protease accelerated the metabolism of postmortem muscle. It can be concluded that trypsin-like protease migrated from the hepatopancreas to the muscle tissue, degraded myofibrillar protein, deteriorated the muscle texture, and eventually leaded to the softening of white shrimp.