ABSTRACT
@#Aims:In this study, measurement of colour changes during solid state fermentation (SSF) are presented as one of the potential techniques that can be used to describe growth, complementary to other biomass estimation, such as weight of fungus, spores concentration, organic matter loss, glucosamine and enzyme activity, which is directly related to growth.Methodology and results: In this study, fungal fermentation of Aspergillus awamoriand A.oryzaeare carried out on complex heterogeneous solid media; wheat bran, soybean hull and rapeseed meal, which are constituted of various soluble and insoluble solid particles. Fermented mass was extracted using distilled water to obtain a cell free extract, which canbe determined quickly and accurately using UV-Visible spectrophotometermeasured absorbance at 300 nm. The results showed a significant correlation between colours produced from fungal SSF and the concentration of spores, weight of fungus cells, organic matter loss, glucosamine and enzyme activity. We found that the colour density proportionally increased when the studied parameters mentioned above increased.For the growth of A. awamoriand A. oryzaeon wheat bran, soybean hulls and rapeseed meal, it was confirmed that colour production was directly proportional to fungal growth. In general, colour-based methods seem to be the most promising approach for biomass estimation in SSF.Conclusions, significance and impacts of study:The theory of measuring colour changes in SSF by UV-visible spectroscopy demonstrates that the colour method gives some valuableinformation than just obtaining a visual observation or spore count to describe growth of fungal mycelium in SSF. The ideas obtained from this exercise might provide a quick and convenient method for quality control of fungal growth. The advantages of this method are that the procedure is simple to carry out, it is non-destructive and no special and expensive reagents are required and the process is very cheap.This newapproach is an important complimentation to the existing techniques especially for basic studies.
ABSTRACT
Aims: Previously described as non-favorable-microbial habitat, peat swamp forest has its own features, which are extremely acidic, poor in nutrient, water-logged and anoxic environment where rate of decomposition of plant litters is quiet slow. Interestingly, current research has proven that there is diversity of microbial communities in this ecosystem. The main objective of this study is to isolate bacteria from Pekan peat swamp forest soil that play a role in the decomposition of plant litters through cultivation on different agar-based medium. The success of isolation of bacteria from this neglected habitat could open the opportunity in unleashing the specific role of bacteria in peat swamp plant litter degradation as well as potential biotechnological application of these bacteria in lignocellulose-related industry. Methodology and results: To mimic the peat condition that is low in nutrient and comprised of plant debris, M1 and peat agar supplemented with cellulose, glucose, lignin and xylan were used. Specifically, for the isolation of actinomycetes, dry and wet heat pre-treatments were applied to the soil samples. Then, the samples were cultivated on three different agars which were oatmeal agar as well as M1 and peat agar supplemented with glucose. Enrichment method was applied in the isolation of cellulase-producing bacteria. It was found that higher number of bacteria and actinomycetes were successfully isolated from peat agar, followed by oatmeal agar and M1. In fact, more actinomycetes were isolated from soil that was treated with wet heat pre-treatment compared to dry heat pre-treatment and on peat agar compared to M1 and oatmeal agar. This finding is promising, indicating that the application of peat water in the agar-based medium is useful to mimic the actual environment of peat swamp and increase the possibility to isolate indigenous bacteria. Primary screening of isolates from samples enriched with carboxymethyl cellulose (CMC) showed positive result of decolourisation zone on Azo-CM-Cellulose agar indicating the ability of isolates to degrade cellulose compound. Conclusions, significance and impacts of study: The study indicates the effectiveness of different culture media in successful isolation of bacteria including actinomycetes. Using the enrichment method, bacteria that are able to degrade cellulose compound was successfully isolated even though it is well known that plant litter degradation in the peat swamp environment happens at very slow rates.
Subject(s)
BacteriaABSTRACT
Aims: Research on lignin degradation capability is previously restricted exclusively to fungal enzymes. However, recent studies had successfully revealed several soil bacterial strains that were able to produce ligninolytic enzymes. These bacterial ligninolytic enzymes were claimed to be more specific in catalysing cleavage of certain linkages between phenolic units of lignin polymers as compared to fungal enzymes. The present study focuses on screening for ligninaseproducing bacteria isolated from South East Pahang Peat Swamp Forest (SEPPSF) soil using agar-based assay. Methodology and results: Thirteen isolates used in this study, which were selected based on distinctive colony morphology from our previous isolation work, showed decolourisation zone on Azure B plates screening. The ratio of decolourisation zones were measured to the ratio of the colony size and the biggest ratio was 2.22 by isolate AR1. Only 4 out of the 13 isolates were able to grow on lignin plates. Subsequently, the 4 isolates, AR3, AR8, AR10 and AR13 were tested on M1 agar supplemented with 3 ligninolytic enzyme indicator compounds which were tannic acid (TA), guaiacol and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) respectively. All four isolates showed growth on TA plates while only AR10 showed a clear brown coloration. An Intense reddish-brown colour formation was observed around the colony of isolates AR3 and AR10 on guaiacol plates while none exhibited green coloration around the colonies when tested on ABTS plates. Conclusions, significance and impacts of study: Isolate AR10 that was identified as Serratia sp. was perceived to be a potential ligninase-producer, though in-depth analysis has to be conducted in the future to determine the specific ligninolytic enzyme activities and characteristics. The application of different substrates is essential to investigate the ligninolytic potential and reaction of those bacterial enzymes towards different indicator compounds. This study is a preliminary endeavour concerning potential ligninolytic enzymes from bacteria as biocatalysts in various industrial processes. This is the first report on preliminary study for ligninolytic activities of soil bacteria from SEPPSF soil.
ABSTRACT
Aims: The aims of this study were to isolate and characterize antimicrobial producing bacteria from tropical peat swamp forest soils. Methodology and results: Bacteria isolated from peat soil were screened for antimicrobial properties via agar overlay assay. Broth microdilution was performed using crude-cell free supernatant (CCFS) to determine the minimum inhibitory concentration (MIC). One isolate was selected due to its broad spectrum activity and identified as Burkholderia spp. with a maximum identity of 99% via 16s rRNA gene PCR. This isolate was able to produce antimicrobials that were active against several Gram positive bacteria, Gram negative bacteria and also yeast. The antimicrobial activity of the CCFS was stable at a pH range of 1 to 11, temperatures of −20 °C to 80 °C, and after treatment with several proteolytic enzymes: α-chymotrypsin, proteinase K and trypsin, indicating that the antimicrobial produced might not be proteinaceous in nature. It is possible that the isolate can produce polyketides, a type of antimicrobial compound produced by Burkholderia known to be resistant to proteolytic enzymes. However, further work needs to be done to confirm this. Conclusion, significance and impact of study: The presence of antimicrobial producing bacteria signified that tropical peat swamps are indeed a potential source for antimicrobials to combat infections.