Generic placeholder image

Current Biotechnology


ISSN (Print): 2211-5501
ISSN (Online): 2211-551X

Research Article

Effects of Various Stress Conditions to Enhance Polyhydroxyalkanoates Accumulation in Wickerhamomyces anomalus VIT-NN01

Author(s): Nupur Ojha and Nilanjana Das*

Volume 9, Issue 2, 2020

Page: [143 - 157] Pages: 15

DOI: 10.2174/2211550109999200801015453

Price: $65


Background: Polyhydroxyalkanoates (PHA) are polyesters produced by various microorganisms. The major drawback of PHA, which is restricting its large-scale production in industries, is its high cost of raw materials and lower PHA concentration.

Objective: The present study aimed to enhance the production of PHA from Wickerhamomyces anomalus VIT-NN01 by optimizing various nutrient and environmental stress conditions and its characterization.

Methods: Effect of various stress conditions viz. nitrogen limitation, salinity, chemical mutagens (acridine orange, sodium azide), and physical stresses (UV, Low Electric Current (LEC), Magnetic Field Intensity (MFI), sound waves) were optimized to screen the best strategic growth conditions for the maximum accumulation of PHA in W. anomalus VIT-NN01. Instrumental analysis was done to evaluate the various changes that occurred in the treated cells and extracted PHA.

Results: The maximum PHA content was observed on the effect of sound waves (88.74%), followed by LEC (87.8%) and MFI (85.75%). The morphological changes in length, shape, and size of the treated W. anomalus cells were observed by Transmission Electron Microscopy (TEM). Smooth, porous matrix, and pseudo spherical microstructure of the extracted PHA were observed by scanning electron microscopy and TEM analysis. The extracted polymer was identified as poly(3-hydroxybutyrate-co-3- hydroxyvalerate) [P(3HB-co-3HV)] co-polymer comprised of 51.66% 3HB and 48.33% 3HV monomer units based on gas chromatography-mass spectrophotometer and nuclear magnetic resonance spectroscopic analysis. X-ray diffraction analysis revealed the crystalline nature of the extracted P(3HB-co- 3HV). The degradation and melting temperatures were found to be 275.9 and 113.8°C, respectively, through thermogravimetric and differential scanning calorimetry analysis.

Conclusion: These results supported the potentiality of W. anomalus, which tolerated the stress conditions and enhanced P(3HB-co-3HV) production from 60 to 88.74% and showed the novelty of present work. This is the first report elucidating the importance of physical stress conditions viz. low electric current, magnetic field intensity, and sound waves for the significant enhancement of PHA production in yeast.

Keywords: Wickerhamomyces anomalus VIT-NN01, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), stress conditions, low electric current, magnetic field intensity, sound waves.

Graphical Abstract
Wen Q, Ji Y, Hao Y, Huang L, Chen Z, Sposob M. Effect of sodium chloride on polyhydroxyalkanoate production from food waste fermentation leachate under different organic loading rate. Bioresour Technol 2018; 267: 133-40.
[] [PMID: 30014991]
Ferreira AM, Queirós D, Gagliano MC, Serafim LS, Rossetti S. Polyhydroxyalkanoates-accumulating bacteria isolated from activated sludge acclimatized to hardwood sulphite spent liquor. Ann Microbiol 2016; 66(2): 833-42.
Al Rowaihi IS, Paillier A, Rasul S, et al. Poly(3-hydroxybutyrate) production in an integrated electromicrobial setup: Investigation under stress-inducing conditions. PLoS One 2018; 13(4)e0196079
[] [PMID: 29698424]
Ojha N, Das N. A Statistical approach to optimize the production of Polyhydroxyalkanoates from Wickerhamomyces anomalus VITNN01 using response surface methodology. Int J Biol Macromol 2018; 107(Pt B): 2157-70.
[] [PMID: 29054518]
Das R, Saha NR, Pal A, Chattopadhyay D, Paul AK. Comparative evaluation of physico-chemical characteristics of biopolyesters P(3HB) and P(3HB-co-3HV) produced by endophytic Bacillus cereus RCL 02. Front Biol 2018; 13(4): 297-308.
Bhatia SK, Gurav R, Choi TR, et al. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) production from engineered Ralstonia eutropha using synthetic and anaerobically digested food waste derived volatile fatty acids. Int J Biol Macromol 2019; 133: 1-10.
[] [PMID: 30986452]
Koller M. Chemical and biochemical engineering approaches in manufacturing polyhydroxyalkanoate (PHA) biopolyesters of tailored structure with focus on the diversity of building blocks. Chem Biochem Eng Q 2018; 32: 413-38.
Elmowafy E, Abdal-Hay A, Skouras A, Tiboni M, Casettari L, Guarino V. Polyhydroxyalkanoate (PHA): Applications in drug delivery and tissue engineering. Expert Rev Med Devices 2019; 16(6): 467-82.
[] [PMID: 31058550]
Sasidharan RS, Bhat SG, Chandrasekaran M. Amplification and sequence analysis of phaC gene of polyhydroxybutyrate producing Vibrio azureus BTKB33 isolated from marine sediments. Ann Microbiol 2016; 66(1): 299-306.
Van Thuoc D, Quillaguamán J. Improving culture conditions for poly (3-hydroxybutyrate-co-3-hydroxyvalerate) production by Bacillus sp. ND153, a bacterium isolated from a mangrove forest in Vietnam. Ann Microbiol 2014; 64(3): 991-7.
Favaro L, Basaglia M, Casella S. Improving polyhydroxyalkanoate production from inexpensive carbon sources by genetic approaches: A review. Biofuels Bioprod Biorefin 2019; 13(1): 208-27.
Valle A, Zanardini E, Abbruscato P, et al. Effects of low electric current (LEC) treatment on pure bacterial cultures. J Appl Microbiol 2007; 103(5): 1376-85.
[] [PMID: 17953548]
Ranalli G, Iorizzo M, Lustrato G, Zanardini E, Grazia L. Effects of low electric treatment on yeast microflora. J Appl Microbiol 2002; 93(5): 877-83.
[] [PMID: 12392536]
Chen H, Li X. Effect of static magnetic field on synthesis of polyhydroxyalkanoates from different short-chain fatty acids by activated sludge. Bioresour Technol 2008; 99(13): 5538-44.
[] [PMID: 18068360]
Ataei SA, Fatehi M. Biological effect of magnetic field on the production of polyhydroxyalkanoates. Sustain Environ Res 2015; 25(2): 125-30.
Gu S, Zhang Y, Wu Y. Effects of sound exposure on the growth and intracellular macromolecular synthesis of E. coli k-12. PeerJ 2016; 4e1920
[ PMID: 27077011]
Priji P, Sajith S, Sreedevi S, Unni KN, Kumar S, Benjamin S. Candida tropicalis BPU1 produces polyhydroxybutyrate on raw starchy substrates. Starke 2016; 68: 57-66.
Gao C, Qi Q, Madzak C, Lin CS. Exploring medium-chain-length polyhydroxyalkanoates production in the engineered yeast Yarrowia lipolytica. J Ind Microbiol Biotechnol 2015; 42(9): 1255-62.
[] [PMID: 26153503]
Haddouche R, Poirier Y, Delessert S, et al. Engineering polyhydroxyalkanoate content and monomer composition in the oleaginous yeast Yarrowia lipolytica by modifying the ß-oxidation multifunctional protein. Appl Microbiol Biotechnol 2011; 91(5): 1327-40.
[] [PMID: 21603933]
Sandström AG, Muñoz de Las Heras A, Portugal-Nunes D, Gorwa-Grauslund MF. Engineering of Saccharomyces cerevisiae for the production of poly-3-d-hydroxybutyrate from xylose. AMB Express 2015; 5(1): 14.
[ PMID: 25852991]
Zhang B, Carlson R, Srienc F. Engineering the monomer composition of polyhydroxyalkanoates synthesized in Saccharomyces cerevisiae. Appl Environ Microbiol 2006; 72(1): 536-43.
[] [PMID: 16391089]
Poirier Y, Erard N, Petétot JM. Synthesis of polyhydroxyalkanoate in the peroxisome of Saccharomyces cerevisiae by using intermediates of fatty acid β-oxidation. Appl Environ Microbiol 2001; 67(11): 5254-60.
[] [PMID: 11679353]
Poirier Y, Erard N, MacDonald-Comber Petétot J. Synthesis of polyhydroxyalkanoate in the peroxisome of Pichia pastoris. FEMS Microbiol Lett 2002; 207(1): 97-102.
[] [PMID: 11886758]
Desuoky AM, El-Haleem ABD, Zaki SA, Abuelhamd AT, Amara A, Aboelreesh GMS. Biosynthesis of polyhydroxyalkanoates in wild type yeasts. JASEM 2007; 11(3): 5-10.
Santhanam A, Sasidharan S. Microbial production of polyhydroxyalkanoates (PHA) from Alcaligens sp. and Pseudomonas oleovorans using different carbon sources. Afr J Biotechnol 2010; 9(21): 3144-50.
Sathiyanarayanan G, Saibaba G, Seghal Kiran G, Selvin J. A statistical approach for optimization of polyhydroxybutyrate production by marine Bacillus subtilis MSBN17. Int J Biol Macromol 2013; 59: 170-7.
[] [PMID: 23603079]
Samantaray S, Mallick N. Impact of various stress conditions on poly-β-hydroxybutyrate (PHB) accumulation in Aulosira fertilissima CCC 444. Curr Biotechnol 2015; 4(3): 366-72.
Aravind J, Sangeetha HS. A study on effect of mutagenic agents on polyhydroxyalkanoates (PHA) production. J Microbiol Biotechnol Food Sci 2014; 3(5): 384.
Sangkharak K, Prasertsan P. Nutrient optimization for production of polyhydroxybutyrate from halotolerant photosynthetic bacteria cultivated under aerobic-dark condition. Electron J Biotechnol 2008; 11(3): 83-94.
Masood F, Hasan F, Ahmed S, Hameed A. Biosynthesis and characterization of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) from Bacillus cereus FA11 isolated from TNT-contaminated soil. Ann Microbiol 2012; 62(4): 1377-84.
Reddy SV, Thirumala M, Mahmood SK. Production of PHB and P(3HB-co-3HV) biopolymers by Bacillus megaterium strain OU303A isolated from municipal sewage sludge. World J Microbiol Biotechnol 2009; 25(3): 391-7.
Huang P, Okoshi T, Mizuno S, Hiroe A, Tsuge T. Gas chromatography-mass spectrometry-based monomer composition analysis of medium-chain-length polyhydroxyalkanoates biosynthesized by Pseudomonas spp. Biosci Biotechnol Biochem 2018; 82(9): 1615-23.
[] [PMID: 29804521]
Liu J, Zhao Y, Diao M, et al. Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) production by Rhodospirillum rubrum using a two-step culture strategy. J Chem 2019.
Liu Q, Luo G, Zhou XR, Chen GQ. Biosynthesis of poly(3-hydroxydecanoate) and 3-hydroxydodecanoate dominating polyhydroxyalkanoates by β-oxidation pathway inhibited Pseudomonas putida. Metab Eng 2011; 13(1): 11-7.
[] [PMID: 20971206]
Obruca S, Sedlacek P, Koller M, Kucera D, Pernicova I. Involvement of polyhydroxyalkanoates in stress resistance of microbial cells: Biotechnological consequences and applications. Biotechnol Adv 2018; 36(3): 856-70.
[] [PMID: 29248684]
Wen Q, Chen Z, Tian T, Chen W. Effects of phosphorus and nitrogen limitation on PHA production in activated sludge. J Environ Sci (China) 2010; 22(10): 1602-7.
[] [PMID: 21235192]
Cui YW, Gong XY, Shi YP, Wang ZD. Salinity effect on production of PHA and EPS by Haloferax mediterranei. RSC Advances 2017; 7(84): 53587-95.
Bashir SM, Girdhar M, Rehman H, Mohan A. Polyhydroxybutyrate (PHB) production and mutagenesis of halophile isolates from the East African Rift salt lake. Biosci Biotechnol Res Asia 2014; 11: 1273-82.
Yavuz H, Celebi SS. Effects of magnetic field on activity of activated sludge in wastewater treatment. Enzyme Microb Technol 2000; 26(1): 22-7.
Hattori S, Watanabe M, Endo T, Togii H, Sasaki K. Effects of an external magnetic field on the sedimentation of activated sludge. World J Microb Biot 2001; 17(3): 279-85.
Kulkarni SO, Kanekar PP, Jog JP, Sarnaik SS, Nilegaonkar SS. Production of copolymer, poly (hydroxybutyrate-co-hydroxyvalerate) by Halomonas campisalis MCM B-1027 using agro-wastes. Int J Biol Macromol 2015; 72: 784-9.
[] [PMID: 25277119]
Sato H, Hoshino M, Aoi H, et al. Compositional analysis of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) by pyrolysis-gas chromatography in the presence of organic alkali. J Anal Appl Pyrolysis 2005; 74(1-2): 193-9.
Pradhan S, Borah AJ, Poddar MK, Dikshit PK, Rohidas L, Moholkar VS. Microbial production, ultrasound-assisted extraction and characterization of biopolymer polyhydroxybutyrate (PHB) from terrestrial (P. hysterophorus) and aquatic (E. crassipes) invasive weeds. Bioresour Technol 2017; 242: 304-10.
[] [PMID: 28366692]

Rights & Permissions Print Export Cite as
© 2022 Bentham Science Publishers | Privacy Policy