Vegetables have been believed to exhibit antioxidant activities due to its phenolic content. Thus, this study was carried out to determine the total phenolic content of water and ethanolic extracts of Malaysian traditional vegetables and assess their antioxidant activities. Eight samples of Malaysian traditional vegetables were dried and extracted its phenolic compounds using water and ethanolic solvent. Total phenolic content of the extracts were compared and evaluated using Folin-Ciocalteu and PrussianBlue reagent. The antioxidant activity were assessed using ferric thiocyanate assay and DPPH free radical scavenging assays. Results found that total phenolic content of water extracts ranged from 7.08 to 14.76 mg GAE (Folin-Ciocalteu assay) and 3.50 to7.82 mg GAE (Prussian Blue assay). However, the content of phenolic of ethanolic extracts ranged from 5.21 to 15.86 mg GAE (Folin-Ciocalteu assay), and 1.84 to 11.54 mg GAE (Prussian Blue assay). The highest antioxidant activity was observed in water extracts of Etlingera elatior(75.6%) and ethanolic extracts of Sauropus androgynus(78.1%). Results also found that the best half maximal inhibitory concentration or IC50were demonstrated by water and ethanolic extracts of Sauropus androgynus which demonstrated 0.077 mg/mL and 0.078 mg/mL, respectively. Hence, this study obtained that most of the Malaysian traditional vegetables have a potential source of natural antioxidant. Keywords:Folin-Ciocalteu method, Prussian Blue assay, antioxidant activities, Malaysian vegetables

Due to the issues of species substitution and product mislabelling of beche-de-mer worldwide, this study aimed to identify the species of seven gamat-based beche-de-mer specimens from Kuah, Langkawi Island, Kedah, Malaysia based on ossicle shapes and non-protein-coding 12S mitochondrial rRNA gene sequences. In general, ossicles were well extracted from the specimens. At least eight ossicle shapes were observed i.e. large I-shaped rod, perforated plate, table, C-shaped rod, button, rosette, I-shaped rod and X-shaped rod. Except for button, I-shaped rod and X-shaped rod, the other five ossicle shapes are common in Stichopus horrens. However, the species status could not be resolved at this level due to the presence of uncommon ossicle shapes and the physical features of the specimens could not be used as supporting data as they were different from the live or unprocessed sea cucumber. In this study, 12S mitochondrial rRNA gene sequences were analysed using the Basic Local Alignment Search Tool programme for Nucleotides (blastn), resulting in the species identification of the beche-de-mer specimens as S. horrens, known locally as gamat emas (golden sea cucumber) with 96-99% similarity (an average of 98%). The phylogenetic trees based on the Neighbour-Joining method, Maximum Parsimony method and Maximum Likelihood method indicated that all 12S mitochondrial rRNA gene sequences of the beche-de-mer specimens clustered with the reference samples of S. horrens from Pangkor Laut, Pangkor Island, Perak, Malaysia, supporting the BLASTN results and confirming their species status as S. horrens. Furthermore, 10 partial 12S mitochondrial rRNA gene sequences of the reference samples and the beche-de-mer specimens of S. horrens were registered with the GenBank (Accession No.: KX879628-KX879637). Overall, the findings suggested that the species identification of the beche-de-mer specimens using 12S mitochondrial rRNA gene sequence gave better inference than ossicle-shape identification. The outcomes of this study benefit enforcement agencies in their work of monitoring and overcoming the issues of species substitution and product mislabelling of beche-de-mer or commercial dried sea cucumber in Malaysian markets as well as in global markets. � Universiti Putra Malaysia Press.

The presence of antimicrobial substances at low concentration may act as chemical signals in altering metabolic processes that take place in bacteria. The chemical signals may activate lots of signal transduction that result in the secretion of bioactive proteins by the stimulated bacterial cells. The bioactive proteins production is strongly influenced by certain factors such as pH and nutrient sources. Therefore, the study of environmental stress factors and growth conditions is very important in inducing the production of bioactive proteins by microbes. The main objective of this study is to evaluate the effects of environmental stress factors and growth conditions on the production of bioactive proteins by Bacillus subtilis ATCC21332 in the presence of Cymbopogon flexuosus essential oil. The microbial proteins were produced by fermentation process using Nutrient broth with adjustment of pH media (pH 6, 7 and 8), and supplementation with 1% (w/w) of three different nutrients, including carbon sources (glucose, sucrose and starch), nitrogen sources (urea, casein and gelatin) and inorganic salts (calcium chloride, sodium nitrate, sodium dihydrogen phosphate) separately. The extracellular protein produced by B. subtilis ATCC21332 was isolated and analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to detect appearance of new protein bands. Results showed that new extracellular proteins were synthesized by B. subtilis ATCC21332 after being induced with C. flexuosus essential oil and grown in each media with 1% (w/w) of starch, 1% (w/w) of gelatin, 1% (w/w) of casein, 1% (w/w) of sodium dihydrogen phosphate and 1% (w/w) of calcium chloride with approximate size of 30.43 kDa, 30.66 kDa, 31.92 kDa, 31.35 kDa and 30.80 kDa respectively. Hence, B. subtilis ATCC21332 in the presence of 0.01 MIC C. flexuosus essential oil under different conditions are able to produce or secrete new extracellular proteins. � 2018, Malaysian Society of Applied Biology. All rights reserved.

Kojic acid is a well known as natural whitening agent and widely use in skincare product formulation. However, kojic acid are water-soluble, unstable at high temperature for long term storage and cannot be directly incorporated in oil based skincare products. Therefore, natural-based kojic acid ester was produced to improve the properties via esterification reaction in the presence of immobilized lipase. The synthesized kojic acid ester was purified through crystallization method. Besides, bioassay study of the kojic acid ester and kojic acid performances on tyrosinase activity and free radical scavenging activity were also carried out. The result shows that, kojic acid gave the highest percentage of tyrosinase activity (77.16%) at 25% of ester concentration. While for kojic acid ester, it gave the highest percentage of tyrosinase activity (23.71%) at 50% of ester concentration. Whilst, the study on radical scavenging activity shows that kojic acid also yield higher percentage of radical scavenging activity which was 84.24% than kojic acid ester which gave 67.65% only. � 2017 American Scientific Publishers All rights reserved.

A few studies on the presence of microbes and their association in sea cucumbers can be found to date, especially in Apostichopus japonicus. However, such studies on Malaysian sea cucumbers and in the gastrointestines of the echinoderms are still lacking. Therefore, the aims of this study are to isolate and identify associated bacteria in the gastrointestines of two common species of Malaysian sea cucumbers i.e. Holothuria (Mertensiothuria) leucospilota (Brandt, 1835), the most dominant sea cucumber species in Malaysia, and Stichopus horrens Selenka, 1867, the commercial gamat species. A number of six genera of Gram-positive bacteria representing the order Bacillales and the order Micrococcales i.e. Bacillus, Brevibacillus, Lysinibacillus, Staphylococcus, Dermacoccus and Micrococcus are isolated from the gastrointestines of H. leucospilota, as suggested by the phylogenetic trees of partial 16S rRNA gene sequences. Meanwhile, three genera of Gram-positive bacteria that represent the order Bacillales i.e. Bacillus, Brevibacillus and Lysinibacillus are isolated from the gastrointestines of S. horrens. Interestingly, 60% of the bacterial species have been known to contain commercial potentials mainly as antibiotic producers, and only bacteria with commercial potentials were present in the gastrointestines of S. horrens. In contrast to that, bacteria that could be pathogenic were also present in the gastrointestines of H. leucospilota. The presence of all bacteria that have been known to contain commercial potentials in the gastrointestines of S. horrens and more diverse microbial population in H. leucospilota could be due to the higher level of antimicrobial properties in the gastrointestines of S. horrens. However, experiments on antibacterial properties of the isolates have to be done and those proven to contain commercial potential can be exploited towards the development of industrial applications in Malaysia. � Universiti Putra Malaysia Press.

Holothuria (Mertensiothuria) leucospilota is the most abundant sea cucumber species in Malaysia. This study aimed to identify bacteria isolated from the external and internal body parts of H. leucospilota collected from the coastal water of Pangkor Island, Perak, Malaysia. A total of 26 bacterial samples were isolated using streak plate method from eight body parts of two fresh H. leucospilota specimens and from the surrounding surface sediments and seawater. Identification of the bacterial isolates was based on microscopic examination, 16S rDNA amplification and phylogenetic analysis using the neighbour-joining method. Three genera of bacteria were identified namely Vibrio, Bacillus, and Acinetobacter. The genus Vibrio was found to be the main bacterial group associated with the H. leucospilota specimens from Pangkor Island. � 2018, Malaysian Society of Applied Biology. All rights reserved.

Ragi is a traditional dry starter culture frequently used in many fermented food products in Asian countries. Ragi contains starch that can be degraded by amylases provided by bacteria and other microorganisms during fermentation in producing fermented foods. This study was carried out to isolate bacteria from ragi and to screen for their ability to hydrolyse starch. Seven bacterial colonies were isolated from ragi, with three samples showing starch hydrolysis activity. Two of the starch-degrading bacterial isolates were identified using partial 16S rRNA gene sequencing as Bacillus licheniformis with 97% and 98% similarity, and one sample identified as a lactic acid bacteria, Enterococcus faecium (98%). Isolation and identification of these bacteria from ragi can provide a promising source of amylase that can be further studied and manipulated for the development of starter culture and to improve the quality of traditional ragi-based fermented food products. � 2018, Malaysian Society of Applied Biology. All rights reserved.

The abundance of sea cucumbers (Phylum Echinodermata: Class Holothuroidea) in Malaysian waters has been gradually declining in past decades. Due to the lack of recent studies on the status of sea cucumber populations in Kudat, Sabah, Malaysia, this study was conducted. This study aimed to identify the species of a timun laut morphospecies i.e. Holothuria (Metriatyla) scabra and two gamat morphospecies i.e. Stichopus horrens and Stichopus ocellatus that were collected from Limau Limawan based on ossicle shapes and non-protein-coding 12S mitochondrial rRNA gene sequences. A number of five main ossicle shapes were microscopically identifed without microscopic size measurement i.e. rod, plate, rosette, button and table. However, a number of ossicle shapes for S. horrens and S. ocellatus recorded in the previous studies were not observed in this study and this could be due to the body deformation of the specimens. Interestingly, fve specimens of H. scabra exhibited additional ossicle shapes other than the smooth button and the table. Despite the absence of common ossicles and the presence of additional ossicle shapes, 12S mitochondrial rRNA gene sequences analysed using the Basic Local Alignment Search Tool programme for Nucleotide (blastn) resulted in the species identifcation of the specimens of morphospecies H. scabra and S. horrens as H. scabra and S. horrens; however, the specimen of morphospecies S. ocellatus was identifed only up to the genus level i.e. genus Stichopus, showing the lack of 12S mitochondrial rRNA gene sequences of S. ocellatus in the GenBank until 15 September, 2016. In total, 31 partial sequences of 12S mitochondrial rRNA gene were registered with the GenBank (Accession No.: KX913672-KX913702). The fndings also suggested that species identifcation based on 12S mitochondrial rRNA gene sequence showed better inference than the ossicle shape identifcation. In summary, the three morphospecies were morphologically and genetically verifed as H. scabra, S. horrens and S. ocellatus. Despite the fact that more specimens and more molecular techniques are required to generate better conclusive outcomes, the current fndings give better insight into the importance of morphological and molecular approaches and the present status of the timun laut species and gamat species in Kudat. � Universiti Putra Malaysia Press.

This preliminary study aimed to identify a commercial gamat species, Stichopus horrens Selenka, 1867, and a timun laut species, Holothuria (Mertensiothuria) leucospilota (Brandt, 1835), from Pangkor Island, Perak, Malaysia, employing morphological techniques based on the shape of the ossicles and molecular techniques based on the cytochrome c oxidase I (COI) mitochondrial DNA (mtDNA) gene. In Malaysia, a gamat is defined as a sea cucumber species of the family Stichopodidae with medicinal value, and timun laut refers to non-gamat species. S. horrens is very popular on Pangkor Island as a main ingredient in the traditional production of air gamat and minyak gamat, while H. leucospilota is the most abundant species in Malaysia. In contrast to previous studies, internal body parts (the respiratory tree and gastrointestine) were examined in this study to obtain better inferences based on morphology. The results showed that there were no ossicles present in the gastrointestine of H. leucospilota, and this characteristic is suggested as a unique diagnostic marker for the timun laut species. In addition, the presence of Y-shaped rods in the respiratory tree of S. horrens subsequently supported the potential to use internal body parts to identify the gamat species. Phylogenetic analysis of the COI mtDNA gene of the sea cucumber specimens using the neighbour-joining method and maximum likelihood methods further confirmed the species status of H. leucospilota and S. horrens from Pangkor Island, Perak, Malaysia. The COI mtDNA gene sequences were registered with GenBank, National Center for Biotechnology Information (NCBI), US National Library of Medicine (GenBank accession no.: KC405565-KC405568). Although additional specimens from various localities will be required to produce more conclusive results, the current findings provide better insight into the importance of complementary approaches involving morphological and molecular techniques in the identification of the two Malaysian sea cucumber species. � Penerbit Universiti Sains Malaysia, 2015.

This study aimed to resolve the taxonomic status of a morphologically undetermined sea cucumber species of order Apodida from Malaysia (GenBank accession no.: FJ223867) using partial 16S mitochondrial rRNA gene sequences and subsequently to determine the validity of morphological taxonomy of Holothuria species into its current subgenera. The undetermined species clustered with all taxa of Holothuria in previous study. Phylogenetic analyses using maximum parsimony and Bayesian methods suggest that the undetermined species was genetically closer to Holothuria (Lessonothuria) pardalis and Holothuria (Acanthotrapeza) coluber; and its position in both phylogenetic trees further suggests its status as a Holothuria taxon. Subgenera of Holothuria, Merthensiothuria and Metriatyla are monophyletic with strong bootstrap supports and posterior probabilities of clades, thus strengthening their morphological taxonomies. Nonetheless, the non-monophyly of subgenera of Halodeima, Microthele and Platyperona suggests a requirement for their taxonomic revisions using integrative taxonomy. The status of Holothuria (Halodeima) edulis subgroups in the maximum parsimony and Bayesian trees is indistinct and further taxonomic revisions are necessary. In terms of sister relationship, both phylogenetic trees suggest that subgenus Holothuria is a sister taxon of subgenus Roweothuria while the other sister relationships were unclear due to the undetermined species, paraphyly and polyphyly of a number of subgenera. Further studies with more specimens of genus Holothuria from broader geographical locations and various mtDNA genes along with morphological approaches may facilitate to provide better insights into the molecular phylogeny of subgenera of Holothuria.

Kojic acid is an organic acid that widely used in cosmetic, health care, food and chemical industries. However, kojic acid is not stable at high temperature, for long storage and it has low lipophilic properties. To overcome this problem, a study was conducted to improve its properties by enzymatic reaction. This study was focused on the synthesis of kojic acid monooleate from kojic acid and oleic acid using enzymatic catalysts at optimized conditions. This study was divided into three parts starting from enzymatic synthesis of kojic ester or specifically kojic acid monooleate (KAMO) by using a dual enzymes system that consisted of Novozym 435 and Lipozyme RM IM. Then, optimization of four variables such as screening of dual enzymes ratio, temperature, substrate molar ratio and the reaction time via conventional method of one parameter at-a-time approach and lastly verification of the product using Fourier Transfer Infrared (FTIR) spectroscopy. The optimum conditions for the enzymatic synthesis of KAMO were at the dual enzymes ratio (1:1) of Novozym 435 and Lipozyme RMIM, temperature at 50C, the molar ratio of kojic acid and oleic acid were 4:1 respectively and 24 hours of reaction time. The highest percentage conversion of KAMO achieved at the most optimum conditions was 13.22 %. Finally, kojic ester produced was verified by using FTIR proved the presence of ester bond (C=O) at 1743 cm-1. � 2018 Author(s).

Proteins levels produced by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antimicrobial agents or antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics or natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was focusing on the effect of essential oil from Cymbopogon flexuosus in regulating proteins production by Bacillus subtilis ATCC21332. The Minimum Inhibition Concentration (MIC) of the C. flexuosus essential oil on B. subtilis was determined by using microdilution assay, resulting 1.76mg/ml. The bacteria cells were further exposed to the C. flexuosus essential oil at concentration of 0.01 MIC for 72 h. The proteins were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile showed that a band with approximate size of 30 kDa was appeared for the treated bacteria with C. flexuosus essential oil. Thus, B. subtilis ATCC21332 in stressful condition with the presence of C. flexuosus essential oils at low concentration could induce the protein production. The isolated protein also showed antimicrobial activity against selected Gram-positive and Gram-negative bacteria. Copyright � 2014 Trans Tech Publications Ltd, Switzerland.

This study aimed at documenting species richness and distribution of sea cucumbers (Phylum Echinodermata: Class Holothuroidea) in Malaysia. Sea cucumber collections were conducted from August 2004 until November 2011 from several study sites in Peninsular Malaysia and Sabah. A total of fifty two morphospecies of sea cucumbers from four orders comprising of 12 genera were documented. Thirty eight species were recorded for Sabah, followed by 24 species for Peninsular Malaysia, and 10 species were recorded in both regions. However, nine species required further taxonomic works for their identification. Of the 15 Actinopyga species recorded, 14 species were from Sabah and one species was from Peninsular Malaysia. The order Aspidochirotida in general, and genus Holothuria in particular were the dominant taxa. Holothuria (Mertensiothuria) leucospilota (Brandt, 1835) is the dominant species in Malaysia as it was observed and documented at all collection sites. Future studies on the species richness of sea cucumbers are required in Sarawak marine waters and the molecular phylogeny of the sea cucumbers in order to obtain a better understanding of the evolutionary relationships between the sea cucumbers of Malaysia. � Universiti Putra Malaysia Press.