This action might not be possible to undo. Are you sure you want to continue?
2013 ARCHIVES OF CLINICAL MICROBIOLOGY
Vol. 4 No. 5:1 doi: 10.3823/272
Our Site: http://www.imedpub.com/
PHB production in Azomonas, Acinteobacter and Bacillus species: Isolation, screening and identiﬁcation
Noha Salah Elsayed, Mohammad Aboulwafa*, Khaled Aboshanab, Nadia Hassouna
Department of Microbiology & Immunology, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
* Address: Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University, Al Khalifa Al Maamoun St., Abbassia, Cairo, Egypt. Tel: (202) 22628017. Fax: (202) 24051107.
Background: Biopolymers (polyhydroxyalkanoates; PHA) are the hope for solving problems of synthetic polymers. They have a lot of applications in medicine and industry. However their production cost is still a problem. Our study aimed to solve part of the problems of PHA production. First we chose polyhydroxybutyrate (PHB) as an abundant and very well characterized member in PHA family. Then isolating easy growing bacterial isolates from soil capable of industrial production of polyhydroxybutyrate and estimating minimum time of incubation needed for its maximum production.
Materials and ﬁndings: A total of 251 bacterial isolates were recovered from
various soil samples. Screening for PHA production was done by viable colony staining method using Nile red. A total of 66 isolates appeared to produce PHA. Screening for PHB was done by spectrophotometric analysis where three promising bacterial isolates were obtained. These isolates were fully identiﬁed using microscopical examination, culture characteristics, biochemical reactions and sequencing of 16S ribosomal RNA gene. They were identiﬁed as Acinteobacter baumannii isolate P39, Bacillus cereus isolate P83 and Azomonas macrocytogenes P173. The ﬁnal 16S ribosomal RNA sequences of the respective isolates were assembled, analyzed and submitted into Genbank. Acinteobacter isolate P39 produced 32 % PHB per dry weight after 30 hours of incubation. After 48 hours of incubation Bacillus isolate P83 and Azomonas isolate P173 produced 13 % and 24 % PHB per dry weight, respectively.
Conclusion: Since PHB production had not been extensively studied in both
Acinteobacter and Azomonas, this study valorizes usage of both isolates in PHB production. Further optimization studies will be conducted in the future for maximum PHB production from these three promising isolates.
This article is available from: www.acmicrob.com
© Copyright iMedPub
Keywords: Polyhydroxybutyrate, Acinteobacter baumannii, Azomonas macrocytogenes, Bacillus cereus, biopolymers.
England). Therefore. a lot of research had adopted several cultivation strategies that can simulate these conditions for the efﬁcient production of PHB . Also Protomonas extorquens used methanol as a carbon source and produced 50-60 % of dry weight . and 1 ml trace elements solutions . Chemicals and culture media Different chemicals used in the present study were of high est quality available and obtained mainly from Sigma-Aldrich (Munich. First they are persistent in the environment to the extent that communities are now more sensitive to the impact of discarded plastic on the environment. (Adwic. PHB has a lot of useful properties such as moisture resistance. plastics have some problems. They are widespread not only due to their uses but also due to their stability [2-4]. NaCl (10).3823/272 Our Site: http://www. Egypt) and other local suppliers. Morphologically different colonies were picked up from plates showing separate colonies to be puriﬁed on nutrient agar plate by streaking method. Since there is a need for plastics in our daily life. PHB has a lot of medical applications as developing therapeutic devices for example temporary prostheses. oxygen impermeability and optical purity.2). The aim of our study was screening of different soil samples to get a promising bacterial isolate (s) capable of industrial production of PHB from soil and identify them. it produced 80 % PHB (w/w) of dry cell mass after 60 hours of incubation. It might be noteworthy that PHA has been found both in Eubacteria and Archaebacteria . Three grams of each soil sample were homogenized in 30 ml of MSM broth and incubated for 24 hours at 37 °C in shaker at 200 rpm and dilution series were made in saline and aliquots of 0. While ready-made culture media and media ingredients were obtained from Lab M (Topley house. Moreover.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol. KH2PO4 (1. The plates were incubated for 24-48 hours at 37ºC. 5:1 doi: 10. water insolubility. 2 © Copyright iMedPub . USA). Therefore. Famous example of PHA is 3-polyhydroxybutyrate (PHB). The two most notable features of PHB in medical applications are that it is very biocompatible producing an exceptionally mild foreign body response. it is composed of (g/liter): Na2HPO4 12H2O (10. ferrous ammonium citrate (0. polyhydroxyalkanoates (PHAs) have been drawing much attention because of their similar material properties to conventional plastics and complete biodegradability. Mineral salt medium (MSM) was used for PHB production.1). CaCl2 (0. according to Lee (1995) the degradation product of PHB. Among examples of bacteria studied for PHB production. so there has been an urge for development of biopolymer (biodegradable polymers) materials which must still retain the desired physical and chemical properties of conventional synthetic plastics . recycling and bio. 12].1 ml were taken and surface inoculated using bent glass rod onto R2A agar. Cairo. Recovery of bacterial isolates from soil samples: Bacterial isolates were recovered from soil samples using the method developed by Yilmaz et al. (2006) with minor modiﬁcations [11.01). 4 No. On the other hand. Oxoid (USA) and Difco (Detroit. PHB was the ﬁrst PHA to be discovered and is also the most widely studied and best characterized PHA . Collection of soil samples: Twenty soil samples were collected from a depth of approximately 10 cm below surface (as microbial ﬂora is affected by U. It is accumulated under unfavorable growth conditions such as limitation of nitrogen. Materials and methods Bacterial isolates: A total of 251 isolates were recovered from different soil samples collected from different localities in Egypt. Among the candidates for biodegradable plastics. We aimed that these isolates will pave a way to decrease PHB production cost by its productivity. three-dimensional porous structures as scaffolds for tissue engineering and as controlled/sustained release drug delivery vehicles . MgSO4 7H2O (0. Second plastics originate from ﬁnite resource (petroleum product). phosphorus.06).com/ Introduction Plastics are considered to be an invaluable gift of modern sciences and technology to mankind .imedpub.or photo-degradation. magnesium.V of sunlight) of agricultural ﬁelds and gardens and stored at 4°C till use. NH4Cl (0. Alcaligenes eutrophus. it is highly plausible that implanting PHB in mammalian tissues would not be toxic.2).5). El-Nasr chemical Co. or oxygen in the presence of excess carbon. and that the biodegradation rate is slow . Solutions to plastic waste management include source reduction. incineration. this makes PHB a unique member in PHA family . Germany). PHB are synthesized by both Gram positive and Gram negative bacteria. However. they are used in many applications such as packaging. D (-)-3-hydroxybutyrate has been detected in relatively large amount in human blood plasma. (2005) and Berlanga et al. these solutions added more problems to the existing grave of plastics .
12) at 37 °C for 1 hour with stirring at 160 rpm. PHB determination: The withdrawn sample contained in eppindorff tubes (1. This method depends on conversion of extracted PHB with concentrated sulphuric acid to crotonic acid which was estimated spectrophotometerically at 235 nm. respectively. This was done by transferring a loopful from culture grown on nutrient agar slant to test tube containing 5 ml L. Calibration curve of crotonic acid concentration versus absorbance at 235 nm. Secondary screening was done by spectrophotometric assay [15. 14]. The tube was incubated in shaking water bath at 37 °C for 20 hours at 160 rpm. The pellets were washed with 1ml aliquot each of water. the tested isolates showed high production of PHB were maintained in L.B medium supplemented with 20% glycerol . The solution was left for cooling then it was measured spectrophotometery at 235 nm. The ﬂasks were inoculated with 5 % v/v pre-culture and incubated in shaking incubator at 200 rpm and 37 °C for 48 hours.0419 where y equals to absorbance of measured sample and X represents the crotonic acid concentrations (µg/ml) in the sample solution. Screening for PHB production: The screening was done on positive isolates obtained from primary screening.The agar plates were exposed to UV light to detect accumulation of PHA. ethanol. Figure 1.imedpub. 4 No. All isolates were cultured on MSM agar medium supplemented with Nile red for 4-5 days at 37 °C . The production was carried out in 100 ml Erlenmeyer ﬂask containing 20 ml MSM. The cell pellets were digested with a sodium hypochlorite solution (equivalent to active chlorine 4-6 %. This was done by dissolving 50 mg crotonic acid in 50 ml concentrated sulphuric acid (98 %) and then appropriate dilutions were made with distilled water and the absorbance was measured for the obtained dilutions .com/ Maintenance of recovered isolates: All the recovered isolates were maintained onto nutrient agar slants at 4 °C and sub-cultured every month. About 1 ml hot chloroform was added to extract PHB and After that 10 ml concentrated sulphuric acid (98%) were added to residue and the tubes were kept in a boiling water bath for 15 min. After screening for PHB production. A linear relationship was observed (Figure 1). 5:1 doi: 10. At the end of incubation. density at 20 °C of 1. The insoluble materials containing PHB were collected by centrifugation at 12. Screening for polyhydroxyalkanoates (PHA) production: The screening was done by the viable-colony staining method using Nile red [11.5 ml) previously washed with alcohol followed by hot chloroform (to remove plasticizers) .000 rpm for 5 min. Construction of standard calibration curve: A standard curve was constructed between crotonic acid concentrations and absorbance at 235 nm. © Copyright iMedPub 3 . 16]. The amount of crotonic acid per ml of sample solution was calculated from the standard curve equation: Y=0.B medium.3823/272 Our Site: http://www.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol. The positive isolates gave pinkish ﬂuorescence under UV.The cells were harvested by centrifugation at 12.017 X+0.000 rpm for 10 min. and acetone. volume of 1 ml was taken for PHB determination and 5 ml sample was taken for dry weight determination.
At the center DNA extraction and PCR ampliﬁcation of 16S rRNA gene were carried out for isolates P39 and P173.nih. Cairo. Through Sigma.net/) and the ﬁnal sequence was aligned against sequences on Genbank using Blast (http://blast. 5:1 doi: 10. Sequencing of 16S rRNA gene: Isolates P39 and isolate P173 were sent to Microbiological Resources Center (MIRCEN).imedpub. the PCR products of both isolates were sent and sequenced at Bioneer Company. Biomass determination: A calibration curve was constructed between optical density at 640 nm and dry weight. 20]. The cell pellets were dried at 37 °C for 24 hours.gov/Blast.000 rpm for 5 min and the supernatants were removed carefully by aspiration and the cell pellets were washed once with physiological saline.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol. 4 No. centrifuged at 12. Biochemical reactions were done to promising isolates according to method designed in Bergeys manual .3823/272 Our Site: http://www. Calibration curves between dry cell weight and optical density of the selected test isolates (P39.ncbi. Egypt. P83 and P173). In parallel serial dilutions of bacterial suspension (similar to those used for dry weight calculations) were done and their optical densities were measured at 640 nm (Figure 2). test organisms were cultured as in secondary screening. Ain Shams Universty. Germany. Identiﬁcation of the selected isolates with promising polyhydroxybutyrate productivity: By microscopical examination was done using gram stain and Gram positive isolates were grown on nutrient agar and mannitol salt agar while gram negative isolates were grown on nutrient agar. organism culture and DNA extraction were carried out according to Pospiech and Neumann 1955  and the extracted DNA was sent to Sigma scientiﬁc company for PCR ampliﬁcation of 16S rRNA gene. The values obtained were averaged and the dry cell weights at different dilution levels were calculated. The sequence assembly and DNA analysis was carried out using Staden package program (http://staden. Then the tubes were dried at 37 °C for 24 hours (pre-determined time for constant weight).nlm. Faculty of Agriculture. The percentage of PHB in terms of crotonic acid per dry weight was calculated as follows: Amount of crotonic acid of 1ml culture Dry weight of 1ml culture X100 these tubes containing dried cell pellets.com/ Dry weight determination: 5 ml aliquot contained in preweighed centrifuge tubes was centrifuged and supernatant was carefully removed by aspiration. To establish this calibration curve. the PCR products of isolate P83 was sent and sequenced at GATC Company. Then each tube was weighed. Through MIRCEN. 4 © Copyright iMedPub . Germany. For isolate P83. The dry weight of cells was calculated from subtracting the initial weight from ﬁnal weight of each centrifuge tube from which the cell dry weight of 1 ml culture was determined. MacConky and EMB agar. Sequence analysis: The sequence obtained from company was analyzed to identify bacterial species.cgi) [19. The dry cell weight was calculated by subtracting the weight of empty centrifuge tubes from ﬁnal weight of Figure 2. At the end of incubation period aliquots of 5ml each were taken in pre-weighed centrifuge tubes.sourceforge.
Discussion In this study. �������� For isolate P83. 5:1 doi: 10. 30. This was evidenced by pinkish ﬂuorescence of colonies on MSM-Nile red agar medium under U. Studying time course: The proﬁles of both PHB production and the growth for the three test isolates (P39.nlm.com/ Studying the time course of PHB production: The three isolates were pre-cultured in LB broth at 37 °C for 20 hours then inoculation in main medium (MSM) was done as in screening for PHB. After dilution R2A medium was used as Reasoner et al. The PHB production paralleled growth for isolate P39 while in case of isolate P173 maximum growth preceded maximum PHB productivity (obtained after 48 hours). These species were identiﬁed by microscopical examination. culture characteristics.V (Figure 3). we were able to isolate 3 promising bacterial strains out of 251 isolates capable of producing relatively high concentration of PHB. KC876036 for Acinetobacter baumannii isolate P39 and KC876035 for Bacillus cereus isolate P83. Results Recovery of bacterial isolates from soil samples: A total of 251 different isolates were recovered from 20 different soil samples. Isolation was carried according to Berlanga et al. The full assembled 16S ribosomal RNA sequences of the isolates P39. PHB concentration and dry weight were measured after 6. P83 as Bacillus cereus and P173 as Azomonas macrocytogenes. culture and biochemical reactions were found to be correlated with the results obtained for 16S rRNA sequence.ncbi. For the other two selected isolates. © Copyright iMedPub 5 . Acinetobacter P39 produces 32 % per dry weight PHB after 30 hours. culture and biochemical reactions are similar to those of Bacillus cereus according to Bergyes manual of systematic bacteriology except for the two tests citrate utilization and tyrosine degradation. 48 and 72 hours of incubation in the main culture. For isolate P83 the increase in growth over time was at slow rate while the increase in PHB production was at higher rate reaching its maximum after 48 hours. biochemical reactions and 16S rRNA gene sequencing. Screening for polyhydroxyalkanoates (PHA) production: A total of 66 isolates accumulate HA.imedpub. Therefore these isolates were selected for further studies.3823/272 Our Site: http://www.gov/genbank/) and the submitted sequences were accepted with accession codes KC685000 for Azomonas macrocytogenes isolate P173. 4 No.nih. (2006) but with some modiﬁcation in the medium used for isolation of microorganism. Fluorescence of PHA producing isolates on MSM. Taken together (microscopical. while bacillus produces PHB up to 13 % per dry weight and Azomonas produces 24 % PHB per dry weight after 48 hours of incubation. the obtained microscopical. culture and biochemical results plus sequence analysis) the test isolate P83 could be identiﬁed as Bacillus cereus. 99 % homology of P83 to Bacillus cereus and 90 % homology of P173 to Azomonas macrocytogenes. Therefore the isolate P39 was identiﬁed as Acinetobacter baumannii isolate P39 and isolate P173 was identiﬁed as Azomonas macrocytogenes isolate P173. P83 and P173 were submitted to the nucleotide Genbank database (http://www. Their identity was P39 as Acinetobacter baumannii. The percentages of PHB production per dry weight (speciﬁc productivity) by the test isolates. 24. Screening for PHB production: A total of 52 producing isolates (20 were Gram positive and 32 were Gram negative) gave positive results for PHB production. First soil was homogenized in MSM broth to provide bacteria with minerals essential for growth. 1985 stated Figure 3. The obtained ﬁnal 16S ribosomal RNA sequences showed 100 % homology of P39 to Acinetobacter baumannii.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol. One Gram positive isolate coded P83 and two gram negative isolates coded P39 and P173 produced maximum percentage of PHB production per dry weight when compared with other tested isolates within the same Gram category. P83 and P173) at different time intervals are shown (Figure 4). the obtained microscopical. Identiﬁcation of bacterial species: Preliminary identiﬁcation was carried out using culture media and biochemical reactions.
imedpub. P. Law and Slepecky (1960) mentioned that PHB cannot be used as standard because they are contaminated with esters and probably with water and it necessities puriﬁcation. This experiment was done in triplicate. The ﬁrst group. and an excess of the carbon sources.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol. The second group accumulates PHA during the growth phase. accumulating PHA during the stationary phase. 4 No. Karr et al. for example. We chose PHB as an example of PHA. Mg and oxygen. 5:1 doi: 10. The medium used in production of PHB is MSM. This method is based on the direct inclusion of the lipophilic dyes Nile red in the agar medium such that the growth of the cells is not affected and the occurrence of PHAs in the colonies can be directly monitored. screening for PHB was done by spectrophotometric assay. requires limitation of N. The screening for PHA was done according to Spiekermann et al. This method gives a powerful discrimination between positive isolates accumulating PHA and negative isolates. Bacillus isolate P83 (b) and Azomonas isolate P173 (c). Crotonic acid was used as standard instead of PHB. Time course of PHB production and biomass in the three selected isolates ( Acinteobacter isolate P39 (a). It is abundant in nature and has a lot of good properties distinguishing it from other PHA . (1999).com/ Figure 4. At the end of incubation in MSM. Since PHB are synthesized and accumulated under unfavorable growth condition in the presence of ex- cess carbon source . Generally bacteria able to synthesize PHA can be divided into two groups. 1983 stated that crotonic acid better used as a standard as purity of PHB differs with © Copyright iMedPub 6 . It is important to develop cultivation strategies that simulate these conditions and allow efﬁcient production of PHB. that R2A medium allow colonies to develop slowly and large enough to be counted easily so it is best used for isolation of bacteria based on morphological differences . It is designed to stimulate conditions of PHB production.3823/272 Our Site: http://www.
com/ the puriﬁcation method used so crotonic acid was used as standard. which makes it possible to identify bacteria to the genus or species level by comparison with databases in the public domain . Dilution of samples with distilled water occurred to decrease viscosity of sulphuric acid . They produced a relatively high concentration of PHB in comparison to their morphological group. (2011) stated that Bacillus subtilius produces 23 µg/ml after 24 hours . isolate P83 among Gram positive isolates and isolates P39 and P173 among Gram negative isolates. we picked up three isolates. This will include both physiological and environmental factors that inﬂuence maximum PHB production by the respective isolates. Further studies will be conducted to optimize PHB production by the three selected isolates.imedpub.mycoides 69 % PHB after 24 hours . 16S rRNA gene sequencing was done as ribosomal 16S exists universally among bacteria and includes regions with species speciﬁc variability. 4 No.3823/272 Our Site: http://www. Nazan et al. PHB production was studied deeply over the last years. Identiﬁcation of the selected isolated were done by culturing them on different media after knowing their Gram behavior. After screening. © Copyright iMedPub 7 . Azotobacter species are well known for industrial production of PHB . Afterwards the polymer concentration decreases due to the bacterial consumption. Gram positive isolates lack lipolysaccarides so its PHB is used in biomedical applications so Bacillus is the best candidate for industrial production . (2002) stated that B.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol.subtilius produces 18 % PHB after 45 hours and B. It makes use of its carbon and energy reserve for drastic conditions. Mostly the isolates produced our polymer in their stationary phase. Time course of PHB production in the three selected isolates was studied. PHB accumulation was stated in other nitrogen ﬁxing bacteria as Azotobacter. sodium hypochlorite was used to digest cells at 37 °C. Acinetobacter was not studied extensively in literature. For efﬁcient recovery of polymer.7 % after 45 hours . Bacillus cereus and Azomonas macrocytogenes. According to Azomonas species. megatiurum produces 14. Biochemical reactions were done afterwards to assure identiﬁcation of bacterial species. It started its production in growth but maximum production in stationary phase. However PHB production in Bacillus was studied to a great extent. Borah et al. Gavin et al (1992) pointed 7 % PHB production in Acinteobacter after 24 hours . (2003) stated that B. According to our study we concluded PHB production in the three species Acinetobacter baumannii. Being biodegradable and from renewable sources attracted a lot of scientists. 5:1 doi: 10. The residue of hypochlorite digestion was washed with distilled water to remove soluble salts and with acetone and alcohol to remove non-PHB lipid. This eliminates the need for puriﬁcation as an extra step. Yuksekdag et al.
15. GE. 3. A Rapid Spectrophotometric Assay of Alpha. RC. U. K. 8. A. Ramsay. K. Adv Biochem Eng. Anderson. BH. PC. Pillai. Kafkas Univ Vet Fak Derg.. 54: 450-472. Boyette. 9. Production of Poly (hydroxyalkanoic acid). P. Rees. 10: 4907-4919. JP. pathophysiology.3823/272 Our Site: http://www. 4 No. SK. Production of poly-bhydroxybutyrate in Acinetobacter spp. Chang. Higuita. 16. PA. A New Medium for the Enumeration and Subculture of Bacteria from Potable Water. Chem.. Geldreick. B. C. Thirumala. EA. J of Med Microbiol 2009.. 21: 565-566. CKS. Appl Enviromen Microbiol 1984. S.... GN. Lenz..β hydroxybutyrate in Bacillus mycoides RLJ B-017. Batielle.com/ References 1. Cardona. 11: 217-218. Bioresour 2013. Woo.. 9: 95-102. 52: 28-58.A Review on Recent Trends and Emerging Perspectives. Reddy. B. 2006. HN. PS. Biodegradable Polymers. Holmes. Int Microbiol. Wu. 22. 27. and genomics studies are also welcome. Y. C. Biotechnol Bioeng 1995. Recent advances in microbial polyhydroxyalkanoates. Poirier. epidemiological. Aslım. M.. 92: 776-783. CA. 26. Teng. RA. Luckachan. Bacterial and other biological systems for polyester production. B.. KV. R. 2009. Guerrero. M. Production of Poly-(P-Hydroxybutyric-Co-3-Hydroxyvaleric Acids. Biotechnol 2005. Occurrence. T. Valorization of glycerol through the production of biopolymers: The PHB case using Bacillus megaterium. 5:1 doi: 10.. May. R. 32: 1697-1699. Somerville. 16S rRNA sequencing in routine bacterial identiﬁcation: A 30-month experiment. Tibtech 1998. A.. RC. Khanna. 1990. Production of polyhydroxyalkanoates a family of biodegradable plastics and elastomers. A. Morgan. Adv in Biochem Eng. C. Vibrio harveyi. ✓ Clinical microbiology relevant inmmunology. 28.. A. M. JL. The inﬂuence of nutritional and environmental conditions on the accumulation of poly.imedpub.... H. in bacteria and plants. 2006. Ashok. J Appl Microbiol. S. Brandl.... JW. preventive measures. Bacterial Polyhydroxyalkanoates. 1990. 10.. JA. 19.and Regulation by the lux Autoinducer.. SY. Bergeys’ Mannual of Systemic Bacteriology (ed.com ✓ Archives of Clinical Microbiology (ACMicrob) is a new peer-reviewed. A versatile quick-prep of genomic DNA from Gram-positive bacteria..β -Hydroxybutyrate (PHB) Production by Bacillus subtilis ATCC 6633 Under Diff erent Conditions. World J. 95: 1798-1803. P. 1995. Sidal. Join Now! http://medicalia. ✓ ACMicrob is an open access journal with rapid publication of articles in all fields and areas of microbiology and infectious diseases. Berkeley.. 19: 637-676. Bayly. ZN.. Gross. treatment. 269: 20785-20790. Appl Microbiol Biotechnol 1992.. 21. Berlanga. N.. J Microbiol Meth. K. Biosynthesis of Poly-3hydroxybutyrate in the Luminescent Bacterium. JC. H. W. Investigation of Poly.. BA. MC. Reasoner. 18. Bacteriol.. Williams and Wilkens) Genus Bacillus. Flandrois. Lee. Posada. J Microbiol Meth. Beta-Unsaturated Acids and Beta-Hydroxy Acids. Dube. 171: 73-80. Afr J of Biotechnol 2004. Lindsay. Stockdale. 1990. and Biodegradable Polyesters. 2011. Process Biochem. Beyatlı.com/ Submit your manuscript here: http://www. Metabolism.. J. JG... JN.. Baumeister. Microbiol. 2002. Lomaliza.. FP. Effect of carbon and nitrogen sources and incubation times on poly-beta-hydroxybutyrate (PHB) synthesis by Bacillus subtilis 25 and Bacillus megaterium 12. Medicalia. SY. SR. Yüksekdag.imedpub. Slepecky. Fernández-Borrell. Metabolic Role. 58: 1030-1036. ✓ ACMicrob covers all aspects of basic and clinical microbiology relevant to infectious diseases including current research on diagnosis. 16: 419-427. H. JK. EA. J. 1961. Microbiol Rev. 13. Guidelines for interpretation of 16S rRNA gene sequence-based results for identiﬁcation of medically important aerobic Gram-positive bacteria. 2005. 2011. viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Naranjo. Y. 41: 77-93.. Phys Technol 1986.. Vasiliadis. Follow us: Publish with iMedPub http://www.acmicrob. Meighen. Spiekermann. MT. Applications of PHB. Mod Plastics 1992. 25. management.. et al. SV. Arch Microbiol.ning. Nazan. 1968. Rapid spectroﬂuorometric screening of poly-hydroxyalkanoate-producing bacteria from microbial mats.isolated from activated sludge. 24... Determination of polyβ -hydroxybutyrate (PHB) production by some Bacillus spp. 17. Fuller. Leung. review their cases and share clinical knowledge. 29. and Industrial Uses of Bacterial Polyhydroxyalkanoates. Dawes. JM. and methodology. Biotechnol 1995.. Sperry. 5. Appl Environ Microbiol. Tse. 69: 64-69. N. genetics. 6. Plastics from Bacteria and for Bacteria: Poly(β -Hydroxyalkanoates) as Natural. 16: 32. 133: 38-44. 17: 173-176. vaccination. 23. Truely degradable resins are now truely commercial. J. Goforth. 4.. D.. Neumann. Chaverie. 1986. Sun.iMedPub Journals 2013 ARCHIVES OF CLINICAL MICROBIOLOGY Vol. B. Montero. EE. Cao. Kalscheuer.. D. Anal Chem. F. Singh.. 11. 7. 14. DW. Thakur. 67: 574-581. RW.. Trends Genet. 3: 63-66. Pospiech. Teng. Steinbüchel. G. et al..org Where Doctors exchange clinical experiences. 2. Parmar. Afr j of Biotechnol. 20. EA. Dawes.N-(3-Hydroxybutanoyl)homoserine Lactone. Mercan. A sensitive. Ribbons. 38: 734-737. Mignard. 1998. 40: 607-619.com 8 © Copyright iMedPub . C.. Law. R. Comparison of the Biolog OmniLog Identiﬁcation System and 16S ribosomal RNA gene sequencing for accuracy in identiﬁcation of atypical bacteria of clinical origin. B.a microbially produced biodegradable thermoplastic.. international journal with world famous scientist on the editorial board. 13: 142-150.. Rehm. 49: 1-14. Füchtenbusch.. Borah... Nawrath. 79: 336343. Biotechnol 1995. 49 (1): 1-7. J Polym and the Environ 2011.. Occurrence of PolyHydroxybutyrate in the Azotobacteriaceae. Biocompatible.. Isolation and characterization of two novel polyhydroxybutyrate (PHB)-producing bacteria. Mahmood.. Steinbüchel. 56: 2093-2098. 12. DJ. Nigam. RCW. Lee. You can also access lots of medical publications for free. Greene. The j of Biol. P. 2: 1105-1139. Claus. 1994.
This action might not be possible to undo. Are you sure you want to continue?
We've moved you to where you read on your other device.
Get the full title to continue reading from where you left off, or restart the preview.