INFLUENCE OF DIFFERENT NITROGEN SOURCES ON GROWTH AND PHB PRODUCTION OF BACTERIAL ISOLATES
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Polyhydroxybutyrate (PHB) is ne of the polyhydroxyalkanoates (PHAs) which has biodegradable and biocompatible properties. Samples were collected form groundnut farm garden in Tsaragi at share in Kwara State. A total of twenty one (21) bacteria were isolated form the soil sample in which bacillus sp were 12 in number while the % frequency of occurrence is (54.5%). Veillococcus sp is 3 and the % frequency of occurrence is (9.1%)s, enterococcus sp, corynebacterium sp, Yersinia Pesis, Lactobacillus fermenta and citrobacter sp were all 1 in number and there % frequency of occurrence is (4.5%) each. The isolates were screened for PHB production using Sudan III stain as well as submerged fermentation. Four (4) of the best PHB producing bacterial were selected for PHBs production.
The fermentation pattern of PHB production was studies using Nitrogen source. NH4Cl (1.560a) was found to be the best effect of PHB growth producing bacteria i.e. (citrobacter sp) and KNO3 (1.899a) the highest for PHB production.
The studies carried out shown that nitrogen sources influence the production of PHB i.e. (citrobacter sp).
TABLE OF CONTENT
Table of content
1.1 Aims and objectives
1.2 Statement of problem
2.0 Literature review
3.0 Materials and method
3.1 Preparation of media
3.2 Isolation of bacterial from soil samples
3.3 Maintenance of culture
3.4 Identification of bacterial isolates
3.4.1 Morphological characterization
3.4.2 Biochemical characterization
3.5 Screening o isolate for PHB production using staining techniques.
3.6 PHB production
3.7 PHB extraction
4.0 Result and discussion
Conclusion and appendix
Polyhydroxybutyrate (PHB) is one of the polyhydroxyalkanoates (PHAs) which has biodegradable and biocompatible properties. They are adopted in the biomed filed, in for example, medical in plants and drug delivery carriers (Keshavarz and Roy, (2010).
Polyhydroxybutyrate was discovered and identified as a granular component in bacterial cells. PHB can grow in a wide variety of natural environmental and is the reserve polymer (intracellular granules) forum in many types of bacteria in nature e.g. in soil, sea water, sewage sludge or compost.
Polyhydroxybutyrate (PHAs) can be classified into two groups depending on the numbe3r of carbon atoms in the monomer unit: short-chain-length. (Scl-PHA) 3-5 carbon atom containing monomer and medium chain-length (MclPHAs) 6-14 carbon atom containing monomers (Steribuchel and Valentin, 1995).
The discovery if a polyester by Smet et al, (1983) was the first report of accumulation of medium-chain-length (MCl-PHAs), i constituents of 6-14 carbon atoms in axenic culture (Smet et al., 1983).
Currently, more than 140 hydroxyalkanoic acid have been identified as constituent of polyhydroxyalkanoates (PHAs) representing a versatile class of microbial polymer (Sterinbuchel and Valentin, 1995). Besides linear and branched 3-40,5- ad 6-hydroxyalkaniates, various functionalized PHAs constituents such as polyhydroxyalkanoates (PHAs) containing halogenated or aromatic side chains have been described (Abe eta l., 1990).
The physical properties of the homopolymer of hydroxybutyrate (PHB) are similar to those of poly propylene, for example regarding melting point, crystrallinity, glass transition temperature etc, and represent a stiff and brittle material (itocking and Marchessault,1994). The properties are improved in copolymers containing up to 25 mol% 3-hydroxy valerate (3HV), where toughness and flexibility are increased and the decrease in crystrallinity and melting point advance the melt-processing of the polymer without being degraded(Marchessault and 4u,2002)
Therefore, the poly (3HB-CO-3HV) copolymer gained industrial interest and in the late 1980s, commercialization under the trade name Biopol was initiated (Holmer etal., 1984). Biopol can be processed to useful materials by various processes such, as extrusion, injection molding, fiber spinning, coating or foaming. Because of the versatile applications as themoplastie biopolymers, polyhydroxy allcanoates (PHAs) can also be used for natural fiber composites or as binder in paints and for various application in medicide and pharmary such as tissue engineering (Kessler et al., 1999)
Polyhydroxylbutyrate(PHB) is a commonly found substance and readily biodegradable are robically and anaerobically. Microbes can use polyhydroxylbutyrate (PHB) exists in the cytoplasmic fluid in the from of crystalline granule about 0.5 Hm in diamete. Beta-hydroxybutyrate is connected by ester linkage and from polyhydroxybutyrate (Andreson and Dawes, 1990). This can be extracted from the cells as native granule or by solvent extraction (Doi and lee, 1990) and processed in the way as polypropylene. S
These biopolymers were also found to increase the resistance of bacteria (Tombolini and Nuti, 1989). Lemoigne, Grelet and croson (1958) draw attention to the different amount of poly-beta – hydroxybutyrate obtain by Bacterim Megaterium on different media and maraca and Wilkinson (1958) showed that more of the substance was formed as the glucose concentration of the growth medium was increased; the subsequest depletion of the product during the later stage of growth suggested a storage function. They also showed that organism rich. In poly-beta-hydroxyrate had a slower rate of autolysis than organisms poor in poly-beta-hydroxybutrate. It is probable; therefore, that poly-beta-hydroxybutyrate act as a reserve carbon and energy source – Tinelli(1955) found that the major part of the material was metabolized at sporulation and deduced that the two processes were intimately connected (Kato et al.,1992)
The bacterium capable of producing polyhydroxybutyrate has been identified in more than twenty (20) bacterial genera, including Azatobacter, Bacillus< Beijernickia, Pseudomonas. Alcaligenes, Rhizobium and Rhodospirillum (Sudesh et al., 2000). many researchers have explained that soil bacteria generally produce polyhydroxybutrate,polyhydroxybutyrate PHB) production increases if ambient conditions (ph, temperature, nutrients) are made available (Hanzlikova et al., 1985).
Pollyhydroxyalkanoate is one such biodegradable microbial polmer which is accumulated in bacteria as intracellular storage and limited nitrogen source (Anderson and Dawes, 1990).
The polymer is known to occur as intracellular granules in several genera of micro-organisms. The granules are syuthesized by prokaryotes acids, sugars and other carbon sources (Atking and kennedy, 1985). Polyhydroxybutrate is insoluble in water, resistant to ultraviolet radiation and is impermeable to oxygen and is very much suitable for use as food packaging material this polymer is readily degraded in the soil and sewage and can be processed using the extrusion technology that is currently used in making polyethylene or polypropylene films(Byrom, 1987). The polyhydroxyalkanoate content and its composition are influenced mainly by the strain of the micro-organisms (Halami, 2008). To achieve a cost effective polyhydroxyalkanoate production, the availability of an efficient bacterial strain is a prereguisite and is a focus of interest for many investigation (Green spare et al., 1985).
The polyhydroxybutyrate production using inexpensive carbon sources in the form of starch by the indigenous strain can be advantageous as the complex starch substrates can be used directly without involvement of any hydrolysis step (itahn et al., 1995).
Polyhydroxybutyrate (PHB) is used in food packaging, plastic films, surgical sutures (stitching of the edges of a wound or incision), contrived drug deliver e.t.c (kim et al., 1994). This bioplastic has many applications in bone plates, nails,serews and in the treatment of osteomyclitics (Itanzlikova et al., 1985).
- AIMS AND OBJECTIVES OF THE RESEARCH
Isolation and biochemical identification of polyhydroxybutyrate (PHB) producing bacteria from soil.
- Screaming of polyhydroxybutyrate (PHB) producing bacteria isolated from soil.
- Polyhydrobutyrate (PHB) Production.
- Determination of effect of different nitrogen sources on growth and polyhydroxybutyrate (PHB) production,
1.2 STATEMENT OF PROBLEM
The accumulation of these non-degradable plastic in the environment is a menacing (treating) draw back increasing day by day. Oil based polymers take many years to degrade which poses an environment problem in some areas, and causes deleterious effects to wild life. To overcome this, production of environmental friendly plastic have been discovered.
The continuous exhaustion of fossil fuels led to the research for the production of bio-degradable plastic from renewable sources the production of biodegradable polymers from renewable resources is the need of the hour, in the face of these ecological facts.
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