Tempoyak is a functional Malaysian food (an acid-fermented condiment) which is produced from the pulp of the durian (Durio zibethinus) fruit. The current study aimed to isolate and identify potential exopolysaccharide (EPS)-producing Lactobacillus strains from tempoyak for potential use as probiotics. Seven isolates (DUR2, DUR4, DUR5, DUR8, DUR12, DUR18, and DUR20) out of 44 were able to produce EPS, and exhibited resistance to acid and bile salt compared to the reference strains Lactobacillus rhmnosus (ATCC53103) and L. plantarum (ATCC8014). The seven isolated strains belonged to five different species—L. plantarum, L. fermentum, L. crispatus, L. reuteri, and L. pentosus—which were identified using API 50 CHL and 16S rRNA gene sequences (Polymerase chain reaction, PCR – based). The seven strains displayed different ability to produce EPS (100–850 mg/L). Isolates exhibited a high survivability to acid (pH 3.0), bile salts (0.3%), and gastrointestinal tract model (<70%). Results showed that the auto-aggregation and cell surface hydrophobicity ranged from 39.98% to 60.09% and 50.80% to 80.53%, respectively, whereas, the highest co-aggregation value (66.44%) was observed by L. fermentum (DUR8) with Pseudomonas aeruginosa. The isolates showed good inhibitory activity against tested pathogens, high antioxidant activity (32.29% to 73.36%), and good ability to reduce cholesterol (22.55% to 75.15%). Thus, the seven tested strains have value as probiotics.

This study aims to evaluate the probiotic properties of Lactobacillus strains producing exopolysaccharides (EPS) isolated from different Malaysian fermented foods. Thirty lactobacilli species were isolated, and six of these (TAP1, PIC7, BU11, BU14, TAP16, and TEMP9) were shown to exhibit probiotic characteristics with the potential to produce EPS. The isolates were evaluated for their tolerance to low pH, bile salts, antimicrobial potential, and cell surface properties. The strains were identified using API50CHL and 16S rRNA gene sequences as Lactobacillus belonged to the species fermentum and plantarum. The Lactobacillus strains exhibited high survivability tolerance to low pH and bile salts (above 90%). All strains exhibited auto-aggregation (33.63–59.63%), co-aggregation (24.37–41.9%), and hydrophobicity (16–65.77%) ability. The strains displayed no transferrable antibiotics and antagonistic activity against pathogenic bacteria. Therefore, it indicates that Lactobacillus strains could act as promising probiotic candidates.

Decreasing heat increment through lowering crude protein (CP) level via supplemental amino acids (AA) have long been suggested as a nutritional practice to alleviate negative heat stress effects on broiler performance. However, there is a considerable body of inconclusive reports on optimisation of such practices, whereby bird performance remains unchanged. The exact mechanism underlying the impaired growth performance in birds fed with reduced-CP diets is not clear yet. Furthermore, adding the environmental temperature factor to the situation may complicate the solution. To date, there is no agreement on the extent of reducing CP level via AA supplementation so that growth performance remains unaffected. Evidence suggests that dietary CP could be reduced safely by 2.3% via essential AA supplementation during later ages, when birds are exposed to an average daily ambient temperature of ≤27.3°C. When Gly was added (a non-essential AA source), the margin of CP reduction could be increased to 5.1% without compromising the growth of broilers subjected to cyclic heat stress. Nonetheless, feeding broilers with a similar Gly-fortified, reduced protein diet failed to support optimal performance under hot and humid tropical climates in 1-21-d-old broilers and had a major impact on growth in broilers reared at 34°C. Regardless of supplemental AA composition or the level of CP reduction, the performance of broilers was negatively affected when birds were subjected to chronic heat stress conditions (≥30°C). These discrepancies can be attributed to a wide range of confounding factors, such as the extent of lowering CP level, types of AA used, age and environmental conditions. Accordingly, the addition of Gly may represent a good approach for reducing dietary CP levels for broilers raised under elevated ambient temperature. Reducing dietary CP is recommended when birds are exposed to moderate but not chronic heat stress conditions.

Penicillium candidum (PCA 1/TT031) synthesizes different types of extracellular proteases. The objective of this study is to optimize polyethylene glycol (PEG)/citrate based on an aqueous two-phase system (ATPS) and Response Surface Methodology (RSM) to purify protease from Penicillium candidum (PCA 1/TT031). The effects of different PEG molecular weights (1500–10,000 g/mol), PEG concentration (9%–20%), concentrations of NaCl (0%–10%) and the citrate buffer (8%–16%) on protease were also studied. The best protease purification could be achieved under the conditions of 9.0% (w/w) PEG 8000, 5.2% NaCl, and 15.9% sodium citrate concentration, which resulted in a one-sided protease partitioning for the bottom phase with a partition coefficient of 0.2, a 6.8-fold protease purification factor, and a yield of 93%. The response surface models displayed a significant (p ≤ 0.05) response which was fit for the variables that were studied as well as a high coefficient of determination (R²). Similarly, the predicted and observed values displayed no significant (p > 0.05) differences. In addition, our enzyme characterization study revealed that Penicillium candidum (PCA 1/TT031) produced a slight neutral protease with a molecular weight between 100 and 140 kDa. The optimal activity of the purified enzyme occurred at a pH of 6.0 and at a temperature of 50 °C. The stability between different pH and temperature ranges along with the effect of chemical metal ions and inhibitors were also studied. Our results reveal that the purified enzyme could be used in the dairy industry such as in accelerated cheese ripening.