Environment improvement through valorization of organic industrial waste by synthesis of poly-β-hydroxybutyrate (PHB) using Synechococcus elongatus

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Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-022-00486-0
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Last Page: 0
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Keywords: Waste valorization, Poly-3-hydroxybutyrate (PHB), Synechococcus elongatus , Sustainable development, Biorefinery


Waste valorization is a technology aimed towards the conversion of waste materials into commercially applicable products like chemicals, materials, and fuels. This technology has revolutionized the field of waste management. The ‘biorefinery’ concept based on biomass-based production techniques is pitched to replace oil-based production technologies. The current research study combines these principles and proposes an organic industrial waste valorization model by biosynthesis of poly-3-hydroxybutyrate (PHB) in cyanobacteria Synechococcus elongatus. Dairy, sugar industry, fruit industry and biodiesel industry waste are used in this study. Our findings show that sugar industry waste is the most productive waste media for PHB synthesis and it yielded 9.84% of the biopolymer. The PHA inclusions are visualized by fluorescent microscopy and the chemical characteristics are studied by FTIR and GC–MS analysis. FTIR spectra reveal five peaks at 1178 cm−1, 1280 cm−1, 1379 cm−1, 1718 cm−1 and 2873 cm−1 corresponding to the characteristic chemical groups of PHB. GC–MS spectra show the peak of methyl ester derivative of 3-hydroxybutyric acid at Rt 2.6 min based on the NIST reference library. It elucidates that the PHB yield is influenced by C:N ratio of the waste sample which provides insights on making the bioplastics production process productive. The current study is significant as it projects an apt substitute for synthetic plastics at a low cost and showcases technology for the management of organically rich industrial waste. The development of such models will lead to environmental improvement and help in achieving the goals of sustainable development.

Waste valorization, Poly-3-hydroxybutyrate (PHB), 
              Synechococcus elongatus
            , Sustainable development, Biorefinery



Ahn J, Jho EH, Kim M, Nam K (2015) Effect of C/N ratio on polyhydroxyalkanoates (PHA) accumulation by Cupriavidus necator and its implication on the use of rice straw hydrolysates. Environ Eng Res 20:246–253

Ahn J, Jho EH, Kim M, Nam K (2016) Increased 3HV concentration in the bacterial production of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) copolymer with acid-digested rice straw waste. J Polym Environ 24:98–103

Albuquerque MGE, Torres CAV, Reis MAM (2010) Polyhydroxyalkanoate (PHA) production by a mixed microbial culture using sugar molasses: effect of the influent substrate concentration on culture selection. Water Res 44:3419–3433

Ali A, Kumar G, Senthilnathan N, Radhakrishnan TP, Sasikala Ch, Ramana ChV (2020) “Sporotan” a new fluorescent stain for identifying cryptic spores of Rhodobacter johrii. J Microbiol Methods 177:106019

Aroncon RAD, Lin CSK, Chan MK, Kwan TH, Luque R (2013) Advances on waste valorization: new horizons for a more sustainable society. Energy Sci Eng 1:53–71

Baei MS, Najafpour GD, Lasemi Z, Tabandeh F, Younesi H, Issazadeh H, Khodabandeh M (2010) Optimization PHAs production from dairy industry wastewater (cheese whey) by Azohydromonas lata DSMZ 1123. Iran J Energy Environ 1:132–136

Ballardo C, Barrena R, Artola A, Sanchez A (2017) A novel strategy for producing compost with enhanced biopesticide properties through solid-state fermentation of biowaste and inoculation with Bacillus thuringiensis. Waste Manag 70:53–58

Beltran-Ramirez F, Orona-Tamayo D, Cornejo-Corona I, Gonzalez-Cervantes J, de Jesus Esparza-Claudio J, Quintana-Rodriguez E (2019). In: Abomohra A (ed) Agro-industrial waste revalorization: the growing biorefinery. Intech Open, London

Bengtsson S, Pisco AR, Johansson P, Lemos PC, Reis MAM (2010) Molecular weight and thermal properties of polyhydroxyalkanoates produced from fermented sugar molasses by open mixed cultures. J Biotechnol 147:172–179

Berwig KH, Baldasso C, Dettmer A (2016) Production and characterization of poly(3-hydroxybutyrate) generated by Alcaligenes latus using lactose and whey after acid protein precipitation process. Bioresour Technol 218:31–37

Bhati R, Mallick N (2012) Production and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) co-polymer by a N2-fixing cyanobacterium, Nostoc muscorum Agardh. J Chem Technol Biotechnol 87:505–512

Bhati R, Mallick N (2016) Carbon dioxide and poultry waste utilization for production of polyhydroxyalkanoate biopolymers by Nostoc muscorum Agardh: a sustainable approach. J Appl Phycol 28:161–168

Bosco F, Chiampo F (2010) Production of polyhydroxyalcanoates (PHAs) using milk whey and dairy wastewater activated sludge production of bioplastics using dairy residues. J Biosci Bioeng 109:418–421

Cavalheiro JMBT, de Almeida MCMD, Grandfills C, da Fonseca MMR (2009) Poly(3-hydroxybutyrate) production by Cupriavidus necator using waste glycerol. Process Biochem 44:509–515

Cesario MT, Raposo RS, de Almeida MCMD, van Keulen F, Ferreira BS, Telo JP, da Fonseca MMR (2014) Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Burkholderia sacchari using wheat straw hydrolysates and gamma-butyrolactone. Int J Biol Macromol 71:59–67

Colombo B, Sciarria TP, Reis M, Scaglia B, Adani F (2016) Polyhydroxyalkanoates (PHAs) production from fermented cheese whey by using a mixed microbial culture. Bioresour Technol 218:692–699

Gahlawat G, Soni S (2017) Valorization of waste glycerol for the production of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) co-polymer by Cupriavidus necator and extraction in a sustainable manner. Bioresour Technol 243:492–501

Godswill AC, Somtochukwu IV (2017) Industrial waste management: brief survey and advice to cottage, small and medium scale industries in Uganda. Int J Adv Acad Res 3:26–43

Gupta A, Verma JP (2015) Sustainable bio-ethanol production from agro residues: a review. Renew Sust Energ Rev 41:550–567

Hongzhang C (2015) Lignocelluose biorefinery engineering—principles and applications. Woodhead Publishing, Cambridge

Jeihanipour A, Bashiri R (2015). In: Karimi K (ed) Perspective of biofuels from wastes, 1st edn. Springer, Cham

Juengert JR, Bresan S, Jendrossek D (2018) Determination of polyhydroxybutyrate (PHB) content in Ralstonia eutropha using gas chromatography and Nile Red staining. Bio-Protoc 8:1–15

Kachrimanidou V, Kopsahelis N, Vlysidis A, Papanikolaou S, Kookos IK, Monje Martinez B, Escrig Rondan MC, Koutinas AA (2016) Downstream separation of poly(hydroxyalkanoates) using crude enzyme consortia produced via solid state fermentation integrated in a biorefinery concept. Food Bioprod Process 100:323–334

Kamilah H, Al-Gheethi A, Yang TA, Sudesh K (2018) The use of palm oil-based waste cooking oil to enhance the production of polyhydroxybutyrate [P(3HB)] by Cupriavidus necator H16 strain. Arab J Sci Eng 43:3453–3463

Karthikeyan L, Venkateshan MS, Krishnan V, Tudor T, Varshini V (2018) The management of hazardous solid waste in India: an overview. Environments 5:1–10

Koch D, Mihalyi B (2018) Assessing the change in environmental impact categories when replacing conventional plastic with bioplastic in chosen application fields. Chem Eng Trans 70:853–858

Koller M, Rodriguez-Contreras A (2015) Techniques for tracing PHA producing organisms and for qualitative and quantitative analysis of intra- and extracellular PHA. Eng Life Sci 15:558–581

Kourmentza C, Costa J, Azevedo Z, Servin C, Grandfils C, De Freitas V, Reis MAM (2018) Burkholderia thailandensis as a microbial cell factory for the bioconversion of used cooking oil to polyhydroxyalkanoates and rhamnolipids. Bioresour Technol 247:829–837

Madkour MH, Heinrich D, Alghamdi MA, Shabbaj II, Steinbüchel A (2013) PHA recovery from biomass. Biomacromol 14:2963–2972

Meixner K, Fritz I, Daffert C, Markl K, Fuchs W, Drosg B (2016) Processing recommendations for using low-solids digestate as nutrient solution for poly-ß-hydroxybutyrate production with Synechocystis salina. J Biotechnol 240:61–67

Mendhulkar VD, Shetye LA (2017) Synthesis of biodegradable polymer polyhydroxyalkanoate (PHA) in cyanobacteria Synechococcus elongates under mixotrophic nitrogen- and phosphate-mediated stress conditions. Ind Biotechnol 13:85–93

Munagala M, Shastri Y (2020) Sustainable valorization of sugar industry waste: status, opportunities, and challenges. Bioresour Technol 303:12292–122302

Nath A, Dixit M, Bandiya A, Chavda S, Desai AJ (2008) Enhanced PHB production and scale up studies using cheese whey in fed batch culture of Methylobacterium sp. ZP24. Bioresour Technol 99:5479–5755

Orona-Tamayo D, Valverde ME, Paredes-Lopez O (2019) Bioactive peptides from selected Latin American food crops—a nutraceutical and molecular approach. Crit Rev Food Sci Nutr 59:1949–1975

Park JBK, Craggs RJ (2010) Wastewater treatment and algal production in high rate algal ponds with carbon dioxide addition. Water Sci Technol 61:633–639

Preethi R, Sasikala P, Aravind J (2012) Microbial production of polyhydroxyalkanoate (PHA) utilizing fruit waste as a substrate. Res Biotechnol 3:61–69

Qiu L, Li J, Li Z, Wang J (2019) Production and characterization of biocontrol fertilizer from brewers spent grain via solid-state fermentation. Sci Rep 9:1–9

Ray S, Prajapati V, Patel K, Trivedi U (2016) Optimization and characterization of PHA from isolate Pannonibacter phragmitetus ERC8 using glycerol waste. Int J Biol Macromol 86:741–749

Reddy MK, Amulya K, Rohit MV, Sarma PN, Venkata Mohan S (2014) Valorization of fatty acid waste for bioplastics production using Bacillus tequilensis: integration with dark-fermentative hydrogen production process. Int J Hydrogen Energy 39:7616–7626

Reddy MV, Mawatari Y, Yajima Y, Satoh K, Mohan SV, Chang YC (2016) Production of poly-3-hydroxybutyrate (P3HB) and poly (3-hydroxybutyrate-co-3-hydroxyvalerate) P(3HB-co-3HV) from synthetic wastewater using Hydrogenophaga palleronii. Bioresour Technol 215:155–162

Reeta D, Anil Kumar PK, Chandrashekar A, Shamala TR (2008) Biosyntheis of polyhydroxyalkanoate copolymers in E. coli using genes from Pseudomonas and Bacillus. Antonie Van Leeuwenhoek 94:207–216

Rivero CP, Hu Y, Kwan TH, Webb C, Theodoropoulos C, Daoud W, Lin CSK (2017). In: Jonathan WC, Tyagi RD, Pandey A (eds) Bioplastics from solid waste, 1st edn. Elsevier B.V., New York

Robak K, Balcerek M (2018) Review of second generation bioethanol production from residual biomass. Food Technol Biotechnol 56:174–187

Sawant SS, Salunke BK, Kim BS (2015) Degradation of corn stover by fungal cellulose cocktail for production of polyhydroxyalkanoates by moderate halophile Paracoccus sp. LL1. Bioresour Technol 194:247–255

Shaikh HM, Pandare KV, Nair G, Varma AJ (2009) Utilization of sugarcane bagasse cellulose for producing cellulose acetates: novel use of residual hemicellulose as plasticizer. Carbohydr Polym 76:23–29

Shea F, Watts CE (1939) Dumas method for organic nitrogen. Ind Eng Chem Anal Ed 11:333–334

Shrivastav A, Mishra SK, Mishra S (2010) Polyhydroxyalkanoate (PHA) synthesis by Spirulina subsalsa from Gujarat coast of India. Int J Biol Macromol 46:255–260

Singh AK, Mallick N (2017) Advances in cyanobacterial polyhydroxyalkanoates production. FEMS Microbiol Lett 364:1–13

Suwannasing W, Imai T, Kaewkannetra P (2015) Cost-effective defined medium for the production of polyhydroxyalkanoates using agricultural raw materials. Bioresour Technol 194:67–74

Taepucharoen K, Tarawat S, Puangcharoen M, Incharoensakdi A, Monshupanee T (2017) Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) under photoautotrophy and heterotrophy by non-heterocystous N2-fixing cyanobacterium. Bioresour Technol 239:523–527

Tsang YF, Kumar V, Samadar P, Yang Y, Lee J, Ok YS, Song H, Kim K, Kwon E, Jeon YJ (2019) Production of bioplastic through food waste valorization. Environ Int 127:625–644

Wang YJ, Hua FL, Tsang YF, Chan SY, Sin SN, Chua H, Yu PHF, Ren NQ (2007) Synthesis of PHAs from waster under various C:N ratios. Bioresour Technol 98:1690–1693

Woertz I, Feffer A, Lundquist T, Nelson Y (2009) Algae grown on dairy and municipal wastewater for simultaneous nutrient removal and lipid production for biofuel feedstock. J Environ Eng 135:1115–1122

Zhu C, Chiu S, Nakas JP, Nomura CT (2013) Bioplastics from waste glycerol derived from biodiesel industry. J Appl Polym 130:1–14



We are thankful to The Director, Institute of Science, Mumbai for his valuable support. We extend our grateful acknowledgment to Director, National Facility for Marine Cyanobacteria (NFMC), Tamil Nadu for making available unialgal strain of Synechococcus elongatus. We like to thank Mahanand Dairy, Indapur Sahakari Sakhar Karkhana Ltd and Muenzer Bharat Pvt. Ltd for providing industrial waste for the research purpose. We also acknowledge the assistance of research staff at NIRRH and P.S. Ramanathan Instrumentation Centre (Ruia College) for the PHB characterization studies.

Author Information

Shetye Laukik
Department of Botany, Institute of Science, Mumbai, India
Mendhulkar Vijay D.
Department of Botany, Institute of Science, Mumbai, India