A novel α-amylase was isolated successfully from Glaciozyma antarctica PI12 using DNA walking and reverse transcription-polymerase chain reaction (RT-PCR) methods. The structure of this psychrophilic α-amylase (AmyPI12) from G. antarctica PI12 has yet to be studied in detail. A 3D model of AmyPI12 was built using a homology modelling approach to search for a suitable template and to generate an optimum target-template alignment, followed by model building using MODELLER9.9. Analysis of the AmyPI12 model revealed the presence of binding sites for a conserved calcium ion (CaI), non-conserved calcium ions (CaII and CaIII) and a sodium ion (Na). Compared with its template-the thermostable α-amylase from Bacillus stearothermophilus (BSTA)-the binding of CaII, CaIII and Na ions in AmyPI12 was observed to be looser, which suggests that the low stability of AmyPI12 allows the protein to work at different temperature scales. The AmyPI12 amino acid sequence and model were compared with thermophilic α-amylases from Bacillus species that provided the highest structural similarities with AmyPI12. These comparative studies will enable identification of possible determinants of cold adaptation.

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Kefir drink is a source of probiotic microorganism with remarkable functional and technological properties. The objective of this work is to isolate yeast strains from Malaysian kefir drink and evaluate them for probiotic potentials. In the present study, nine strains of probiotic yeast were isolated from a Malaysian kefir drink and identified according to their 16S rDNA sequences. Furthermore, their probiotic potential was evaluated. The probiotic properties were tested for aspects of antibiotic susceptibility, antimicrobial activity, and gastrointestinal condition tolerance (pH and temperature). Five isolated strains, M3, Y5, Y9, Y11 and A1, showed good tolerance towards low pH condition while three strains, A1, M1, and M3, showed antimicrobial activity against E. coli, P. aeruginosa, and Salmonella sp. Most isolates were resistant to penicillin, streptomycin, and ampicillin, and grew well at human body temperature. The result of this test indicates that the yeast strains isolated from Malaysian kefir drink have excellent potential for use as probiotics in various products. Lastly, kefir milk is one of the excellent source of probiotic yeast strains and could be used as a new yeast probiotic formulation or in food supplements.

In this study astaxanthin production by Phaffia rhodozyma was enhanced by chemical mutation using ethyl methane sulfonate. The mutant produces a higher amount of astaxanthin than the wild yeast strain. In comparison to supercritical fluid technique, high-pressure homogenization is better for extracting astaxanthin from yeast cells. Ultrasonication of dimethyl sulfoxide, hexane, and acetone-treated cells yielded less astaxanthin than β-glucanase enzyme-treated cells. The combination of ultrasonication with β-glucanase enzyme is found to be the most efficient method of extraction among all the tested physical and chemical extraction methods. It gives a maximum yield of 435.71 ± 6.55 µg free astaxanthin per gram of yeast cell mass.

Abstract. Bioactive peptide from casein play a very important role in biological functionalities such as antioxidant and antimicrobial activities. Casein is the main protein that derived from goat milk which consists of alpha (α), beta (β) and kappa (κ) casein. Dietary protein such as casein from animal can provide rich source of bioactive peptide. However, the macromolecular protein such as cow milk can cause allergic response to certain individuals. On the other hand, goat milk have been known for its hypoallergenic and therapeutic properties in human nutrition and health. The purpose of this study is to extract casein from local breed goat milk and identify the molecular characterization of casein for nutraceutical value. The casein was successfully extracted using extraction method. Extraction is a common technique used to separate a desired substance when it is mixed with other components. The average percentage of casein obtained was 24.25%. Then, the casein was analysed by running it in the SDS-Page. The major fraction is β-casein and the minor is α-casein that can be seen between 20kDa and 30kDa respectively. There is no contaminated protein appear in the purified α-amylase. The result obtained in this study indicates that isolated casein from Malaysian goat milk was pure and can be used as bioactive peptide for nutraceutical value.

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