Optimization of Cultural Conditions for the Biosynthesis of Phenylalanine ammonia lyase (PAL) by Bacillus subtilis GCB-31 on agro-industrial wastes

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Research Paper 01/11/2019
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Optimization of Cultural Conditions for the Biosynthesis of Phenylalanine ammonia lyase (PAL) by Bacillus subtilis GCB-31 on agro-industrial wastes

Qasim Shahzad, Sadia Javed, Saqib Mahmood, Arruje Hammed
Int. J. Biosci.15( 5), 233-240, November 2019.
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Phenylalanine ammonia lyase (PAL) is the key enzyme that catalyzes the conversion of L-Phenylalanine to ammonia and trans- cinnamic acid. In phenyl propanoid pathway, PAL is the first enzyme which involved in the biosynthesis of polyphenol compounds such as flavonoids and lignin in plants. The present study was conducted with the main objective to optimize the phenylalanine ammonia lyase (PAL) production process by Bacillus subtilis GCB-31 on agro-industrial wastes as carrier substrates in solid state fermentation. The maximum PAL activity was obtained at 30 oC after 96 h using 1 mL inoculum level. 10g of mixed substrates (Corn Stover+ Corn Cobs+ Banana stalk) was optimized for maximum L-asparaginase production. Agro-industrial wastes, nitrogen sources and carbon sources were also optimized for the maximum production of enzyme (58.77±2.94 U/mL). It was also found that glucose and ammonium sulphate as the best additional carbon and nitrogen sources respectively. It can be inferred from this study that optimization of fermentation process enhanced the production of desired enzyme.


Baskar G, Renganathan S. 2011. Design of experiments and ANN linked genetic algorithm for modeling and optimization of l-asparaginase production by Aspergillus terreus MTCC 1782. Biotechnology and Bioprocess Engineering 16, 50-8. http://doi.org/10.1007/s12257-010-0119-7

Berner M, Krug D, Bihlmaier C, Vente A, Müller R, Bechthold A. 2006. Genes and enzymes involved in caffeic acid biosynthesis in the actinomycete Saccharothrixespanaensis. Journal of bacteriology 188, 2666-73. http://doi.org/10.1128/JB.188.7.2666-2673.2006

Chesters C, Wilding M, Goodall M, Micklefield J. 2012. Thermal bifunctionality of bacterial phenylalanine aminomutase and ammonia lyase enzymes. Angewandte Chemie International Edition 51, 4344-8. https://doi.org/10.1002/anie.201200669

Cui JD, Qiu JQ, Fan XW, Jia SR, Tan ZL. 2014. Biotechnological production and applications of microbial phenylalanine ammonia lyase: a recent review. Critical Reviews in Biotechnology 34, 258-68. https://doi.org/10.3109/07388551.2013.791660

D’Cunha GB, Satyanarayan V, Nair PM. 1996. Stabilization of phenylalanine ammonia lyase containing Rhodotorulaglutinis cells for the continuous synthesis of L-phenylalanine methyl ester/96. Enzyme and Microbial Technology 19, 421-7. https://doi.org/10.1016/S0141-0229(96)00013-0

Dutta JR, Dutta PK, Banerjee R. 2004. Optimization of culture parameters for extracellular protease production from a newly isolated Pseudomonas sp. using response surface and artificial neural network models. Process Biochemistry 39, 2193-8. http://doi.org/10.1007/s13765-014-4194-x

E-A El-Naggar N. 2012. Extracellular production of the oncolytic enzyme, L-asparaginase, by newly isolated Streptomyces sp. strain NEAE-95 as potential microbial cell factories: Optimization of culture conditions using response surface methodology. Current PharmaceuticalBiotechnology 16, 162-78. https://doi.org/10.2174/1389201015666141113123910

Edwinoliver NG, Thirunavukarasu K, Naidu RB, Gowthaman MK, Kambe TN, Kamini NR. 2010. Scale up of a novel tri-substrate fermentation for enhanced production of Aspergillus niger lipase for tallow hydrolysis. Bioresource Technology 101, 6791-6. https://doi.org/10.1016/j.biortech.2010.03.091

Iftikhar T, Niaz M, Nisa Z, Tariq AS, Khalid MN, Jabeen R. 2011. Optimization of cultural conditions for the biosynthesis of lipases by Penicillium chrysogenum (MBL 22) through solid state fermentation. Pakistan Journal of Botany 43, 2201-6.

Javed S, Meraj M, Mahmood S, Hameed A, Naz F, Hassan S, Irfan R. 2017. Biosynthesis of lovastatin using agro-industrial wastes as carrier substrates. Tropical Journal of Pharmaceutical Research 16, 263-9. http://dx.doi.org/10.4314/tjpr.v16i2.

Kong JQ. 2015. Phenylalanine ammonia-lyase, a key component used for phenylpropanoids production by metabolic engineering. RSC Advances 5, 62587-603. http://doi.org/10.1039/C5RA08196C

Lowry OH, Roserbrough N, Farr AL, Randall R 1951. Protein measurement with folin phenol reagent. Journal of Biological Chemistry 193, 265-275.

Moffitt MC, Louie GV, Bowman ME, Pence J, Noel JP, Moore BS. 2007. Discovery of two cyanobacterial phenylalanine ammonia lyases: kinetic and structural characterization. Biochemistry 46, 1004-12. https://doi.org/10.1021/bi061774g

Mukhtar H, Haq I. 2008. Production of alkaline protease by Bacillus subtilis and its application as a depilating agent in leather processing. Pakistan Journal of Botany 40, 1673-9.

Nadeem M, Qazi JI, Baig S, Syed QU. 2007. Studies on commercially important alkaline protease from Bacillus Lichniformis N-2 isolated from decaying organic soil. Turkish Journal of Biochemistry-turkbiyokimyadergisi 32, 171-7. http://doi.org/10.4172/1948-5948.1000103

Panda SK, Mishra SS, Kayitesi E, Ray RC. 2016. Microbial-processing of fruit and vegetable wastes for production of vital enzymes and organic acids: Biotechnology and scopes. Environmental research 1, 161-72. https://doi.org/10.1016/j.envres.2015.12.035

Santiago R, De Armas R, Legaz ME, Vicente C. 2009. Changes in phenolic acids content, phenylalanine ammonia-lyase and peroxidase activities in sugarcane leaves induced by elicitors isolated from Xanthomonasalbilineans. Australasian Plant Pathology 38, 357-65. http://doi.org/10.1071/AP0900908153191/09/040357

Silman RW, Conway HF, Anderson RA, Bagley Production of aflatoxin in corn by a large‐scale Solid‐substrate fermentation process. Biotechnology and Bioengineering 21, 1799-808. http://dx.doi.org/10.1002/bit.260211008

Steel RG, Torrie JH, Dickey DA. 1979. Principles and procedures of statistics. Pages: 400-428. A biometrical approach 3. https://doi.org/10.1002/bimj.19620040313

Varalakshmi V, Raju KJ. 2013. Optimization of l-asparaginase production by Aspergillus terreus MTCC 1782 using bajra seed flour under solid state fermentation. International Journal of Resource Engineering and Technology 2, 121-9. https://doi.org/10.15623/ijret.2013.0209020

Zhu YX, Liao SY, Ye J, Zhang H. 2012. Cloning and characterization of a novel tyrosine ammonia lyase-encoding gene involved in bagremycins biosynthesis in Streptomyces sp. Biotechnology Letters 34, 269–74. https://doi.org/10.1007/s10529-011-0755-9