Fungi Producing Xylanase Using Palm Oil Mill Effluent
1.1 Background of Study
Palm oil processing was carried out using large quantities of water in mills where oil is extracted from the palm fruits. During the extraction of crude palm oil from the fresh fruits, about 50% of the water results in palm oil mill effluent (POME). It is estimated that for 1 tonne of crude palm oil produced, 5 – 7.5 tonnes of water ends up as POME (Ahmad et al., 2003). The solid waste products that result from the milling operation are empty fruit bunches, palm fibre, and palm kernel. In both traditional and modern milling settings, these solid waste products are all put to economically useful purposes such as fuel material and mulch in agriculture. It is the POME that is usually discharged into the environment enzymes are distinct biological polymers that catalyze the chemical reactions and convert substrates to particular products (Haq et al., 2006). Xylanase have gained a unique place in the biotechnological sector due to their potential application in palm oil mill effluent industry (Kirk and Jefferies, 1996), food and feed industry (Bhat, 2000), textile industry (Csiszár et al., 2001) and Biofuel production (Goldschmidt, 2008). Xylanases constitute one of the most important industrial enzymes that depolymerizes xylan molecule into xylose units (Garg et al., 2011). The production of enzymes generally depends on variety of growth parameters like inoculum size, pH value, temperature, inducers, medium additives, aeration, growth and time (Immanuel et al., 2006) and also the enzyme activities depend on the presence of various metal ions as activators and inhibitors (Muhammad et al., 2012). Enzymes have various applications different industries.
The major industrial applications of enzymes are in textile industry for‘bioEndo-1, 4-β-D-xylanases (E.C.184.108.40.206) are hemicellulases accountable for random cleavage of the xylan backbone, thus, are industrially significant. (Zhou et al., 2008). Among the microbial sources, filamentous fungi are particularly interesting since they secrete these enzymes into the medium and have high xylanase activity in contrast to yeasts and bacteria (Krisana et al., 2005). This feature makes fungal xylanases attractive to be used in various industrial processes hence, fungi are highly diverse in nature; they have been recognized as a target for screening to find out the appropriate source of enzymes with constructive and novel characteristics (Bakri et al., 2010).
1.2 Justification of this Study
Xylanases are hydrolases which depolymerise the plant cell wall component-xylan, the second most abundant polysaccharide. They are mainly produced by microorganisms but can also be found in plants, marine algae, protozoans, crustaceans, insects, and snails. Because of their ability to break down xylan, these enzymes especially of microbial origin, have attracted more attention due to their potential role in pulping and bleaching processes, in food and feed industry, textile processes and organic waste treatment. Xylanases are more suitable in paper and pulp industry than lignin degrading enzymes. Owing to the increasing biotechnological importance of thermostable xylanases, many potential thermophilic and hyperthermophilic have been identified. As tolerance to higher pH and temperature are desirable properties of xylanase for effective use in pulp treatment, thermophillic organisms are of special interest as a source of novel thermostable xylanases. But for large scale production of xylanases, reduction of cost is still very challenging. This work will assay the potentiality and activity of xylanolytic fungi on palm oil mill effluent.
1.3 Aim of Study
The major aim of this project is to isolate fungi producing xylanase using palm oil mill effluent.
1.4 Objectives of the Study
The specific objectives are:
- To isolate fungi from palm oil mill effluent using Potato Dextrose Agar
- To identify the morphological characteristics of fungal isolates produced
- To screen for xylanase production by fungal isolates using congo red
- To demonstrate the biodegradation of POME by fungi producing xylanase from palm oil mill effluent.
5.0 Discussion, Conclusion and Recommendation
From the results obtained from the fungal isolates from palm oil mill effluent which are A.niger, A.flavus, F.solani, Rhizopus and Mucor. Three of the organisms were capable of producing xylanase and these organisms include A.niger, A.flavus and F.solani.
Table1 represents the zone of inhibition of the organisms on czapek’s medium. The result shows A.niger to be 9.5±0.3, A.flavus 6.0±1.0 and F.solani5.1±0.1 zone of inhibition respectively. This is in agreement with (Muthezhilan et al., 2007). The data above indicates that, zone of inhibition is not a perfect marker to decide the highest producer, however, might indicate about the selection of isolates (Teather and Wood, 1982; Tseng et al., 2000).
The viable count result showed that there was an increase in the number of cells from day 0 to 8 and then a decrease as the day progressed. This is because the inoculated samples utilized POME as carbon source. The inoculated samples consisted of mineral salt medium, POME and the organism whereas the control consist of just POME and organism without media, the number of cells in the inoculated sample was greater than the control. The values recorded for A.niger from day 0 were 6.1×105 ± 0.07, 4.1×105 ±0.09,6.9×105± 0.08, 4.7×105 ±1.00 and 3.3×105± 1.00 respectively. At the end of bio-degradation (day16th), the growth of cells decreased. The result of A.flavus from day 0 to 16 were recorded as 4.5×105 ± 0.03, 4.2×105 ± 1.00, 5.0×105 ± 0.08 , 3.2×105 ±1.00 and 2.8 ×105 ±0.02,the growth also decreased at day 16th while for F.solani ,the values recorded from day 0 to 16th were 4.8×105 ±0.05, 5.0×105±0.05,4.9×105 ± 0.01,3.7×105 ±0.04 and 3.0×105 ±0.07 respectively. The highest colonies were observed in A.niger and A.flavus which occurred at day 8th which was 6.9×105± 0.08 cfu/ml and 5.0×105 ± 0.08 while the highest observed in F.solani occurred at the 4th day .The reason why the organisms decreased in number could be due to temperature changes, reduction in nutrients or environmental factors. The inbuilt genetic make-up of any organism is accountable for the maximum enzyme production and is largely dependent upon the type of microbial strain as well as on cultural environmental conditions during the growth of the organism (Bajaj and Abbass, 2011).
The physiochemical properties during biodegradation of POME were determined and the results for turbidity were as follows; A.niger from day 4 to 16 1ncluded 0.884±0.085, 0.576±0.006, 0.354±0.03 and 0.706±0.015 respectively, A.flavus for the same period of days included 0.806±0.004, 0.417±0.002,0.348±0.024 and 0.654±0.030 respectively while that of F.solani were 0.554±0.04, 0.317±0.015, 0.318±0.020 and 0.612±0.035 respectively. The turbidity increased with increase in days and therefore day 16th being the most turbid. The turbidity was higher in the sample inoculated with A.niger, A.flavus and F.solani than there various control . This indicates that the inoculated samples were able to grow and utilize the POME as there carbon source. The reason for the increase in turbidity after 4 days till the end of the study may be due to the presence of nitrogen and phosphorus in the mineral salt medium which is necessary for bio-degradative activity(Adesodun and Mbagwu, 2008).And also play a role in overcoming nutrient limitation during the bio-degradative process. Turbidity was highest in A.niger then A.flavus and the least is F.solani with the following values 0.884±0.085, 0.806±0.004, 0.417±0.002, 0.348±0.024 and 0.554±0.04 respectively. This investigation, in accordance with others (Anthony et al., 2003; Ghosh and Nanda, 1994), have showed that the highest xylanase producing strain, with less or no cellulase activity, belong to the genus Aspergillus sp.
After the statistical analysis for the degradability of POME by fungal isolates was done, the result shows that there was no significant difference in the biodegradability of POME by the microbial isolates.
Considering the world market in enzymes, we can see that dramatic changes will be necessary in the future in order to make national enzyme production more competitive. Great hopes are placed on technological advances, but there is also a search for new microorganisms, within the great biodiversity of this planet, that may possess better physiological characteristics in relation to temperature, pH of the medium and adaptability to low-cost substrates, which have been until now hardly exploited. It is therefore concluded that the enzyme (xylanase) degraded by A.niger, A.flavus and F.solani are capable of breaking down POME, and thus industrial wastes such as POME have the potential to produce xylanases.
For further studies on the application of these enzymes in the paper and pulp, food industry, in environmental science, such as bio-fueling, effluent treatment, and agro-waste treatment will require a complete understanding of the functional and genetic significance of the xylanases and further purification of the xylanase. Hence the production of xylanase can be improved by finding more potent fungal strains with improved enxyme expression profiles or higher protein secretion level and should be intensified in other to develop viable enzymatic production technology that employ agro residual waste as substrates.
Fungi Producing Xylanase Using Palm Oil Mill Effluent
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