Halotolerant proteases are known as one of the important groups of enzymes that have been used widely in various industries. However, high production cost of proteases in term of energy used for sterilization and high risk of microbial contamination during fermentation become the obstacles during upstream processing. Moreover, proteases that capable to withstand with harsh conditions of salinity, temperature and pH are required in industry. Therefore, this study presents newly isolated halophilic bacteria producing halotolerant proteases. Halophilic bacteria were isolated from fermented fish sauce. Screening of the bacteria producing halotolerant protease enzymes was carried out by using skim milk salt agar containing 10% NaCl at pH 7 and incubated under aerobic condition at 37oC for 2 days. The selected isolates were identified based on their morphology followed by 16S rDNA gene sequence analysis. 40 colonies of halophiles bacteria were isolated, however, only 20 of them showing proteolytic activity. All of 20 isolates are motile and gram positive bacteria. From 20 isolates, only 6 were chosen for further analysis. B7 showed the highest proteolytic activity compared with others. Results of 16S rDNA gene sequence analysis showed 98% homology to Bacillus amyloliquefaciens subsp. plantarum strain FZB42. Therefore, B7 is identified as Bacillus amyloliquefaciens strain B7.
Growing cells on microcarriers may have overcome the limitation of conventional cell culture system. However, the surface functionality of certain polymeric microcarriers for effective cell attachment and growth remains a challenge. Polycaprolactone (PCL), a biodegradable polymer has received considerable attention due to its good mechanical properties and degradation rate. The drawback is the non-polar hydrocarbon moiety which makes it not readily suitable for cell attachment. This report concerns the modification of PCL microcarrier surface (introduction of functional oxygen groups) using ultraviolet irradiation and ozone (UV/O) system and investigation of the effects of ozone concentration, the amount of PCL and exposure time; where the optimum conditions were found to be at 60,110.52 ppm, 5.5 g PCL and 60 min, respectively. The optimum concentration of carboxyl group (COOH) absorbed on the surface was 1495.92 nmol/g and the amount of gelatin immobilized was 320 ± 0.9 µg/g on UV/O treated microcarriers as compared to the untreated (26.83 ± 3 µg/g) microcarriers. The absorption of functional oxygen groups on the surface and the immobilized gelatin was confirmed with the attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR) and the enhancement of hydrophilicity of the surface was confirmed using water contact angle measurement which decreased (86.93°-49.34°) after UV/O treatment and subsequently after immobilization of gelatin. The attachment and growth kinetics for HaCaT skin keratinocyte cells showed that adhesion occurred much more rapidly for oxidized surfaces and gelatin immobilized surface as compared to untreated PCL.
Astaxanthin is one of the main carotenoid pigments. It has beneficial effects on the immune system of the human body due to its powerful antioxidant properties. The application of this bioactive compound can be found to be significant in the food, pharmaceutical, and cosmetics industries. The aim of this research was to investigate astaxanthin yield from six species of Malaysian shrimp carapace. Six types of shrimp species-Parapenaeopsis sculptili, Metapenaeus lysianassa, Macrobrachium rosenbergii, Metapenaeopsis hardwickii, Penaeus merguiensis, and Penaeus monodon-were used to investigate total carotenoid content and astaxanthin yield. The investigation was carried out using chemical extraction and high-pressure processing (HPP) methods at 210 MPa, for a period of 10 min with a solvent mixture of acetone and methanol (7:3, v/v). HPP was proven to have a significant impact in increasing the total carotenoid content and astaxanthin yield. The highest total carotenoid content and astaxanthin yield is shown to be contained in the Penaeus monodon species. Total carotenoid was increased from 46.95 µg/ml using chemical extraction to 68.26 µg/ml using HPP; yield of astaxanthin was increased from 29.44 µg/gdw using chemical extraction to 59.9744 µg/gdw using HPP. Therefore, comparison between the HPP and chemical extraction methods showed that HPP is more advantageous with higher astaxanthin yield, higher quality, and shorter extraction time.
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