Isolation and Optimization of PHB (Poly-β-hydroxybutyrate) Based Biodegradable Plastics from Chlorella vulgaris Rebecca Robert * and Priya R Iyer Department of Biotechnology, Women’s Christian College, Chennai, Tamil Nadu, India * Corresponding author: Rebecca Robert, Department of Biotechnology, Women’s Christian College, Chennai, Tamil Nadu, India, Tel: +91 044 2827 5926; E-mail: rebeccaobeth@yahoo.com Rec date: February 08, 2018; Acc date: February 15, 2018; Pub date: February 19, 2018 Copyright: © 2018 Robert R, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Poly-β-hydroxybutyrate (PHB) can be used as an effective thermoplastic and has many characteristics similar to those of standard commercial plastics like polypropylene. PHB based plastic substitutes are less flexible than traditional plastics; they are completely biodegradable and leave behind no residue. Algae are used for the production of PHB, for bioplastic production which offers an opportunity in economic efficiency by reduced costs. Chlorella vulgaris PB (1-6) was isolated from different freshwater sources and screened for PHB production using Sudan black B and Nile Blue Stain. The production of PHB was optimized using different media and under various parameters like Aeration; Effect of phosphate and Sodium acetate etc. PHB was extracted using hot chloroform and the amount of PHB produced was estimated by reading the absorbance at 235 nm. Keywords: PHB; Chlorella vulgaris PB (1-6); Biopolymers; Nile blue; Biodegradation Introduction Bioplastics or organic plastics are a form of plastics derived from renewable biomass sources such as vegetable oil, corn, starch, pea starch unlike fossil-fuel plastics derived from petroleum. Biodegradable form of plastic was frst characterized in the mid 1920’s by French researchers. Tis molecule is called Polyhydroxybutyrate (PHB). Many diferent types of bacteria and algae produce it as food storage material [1]. Biodegradable plastics can decompose into carbon-dioxide, methane, water, inorganic compounds or biomass via microbial assimilation. Algae serve as an excellent feedstock for plastic production owing to its many advantages such as high yield and the ability to grow in a range of environments [2]. In this paper, Algae are used for the production of PHB. Industrial utilization of Algae as PHB producers has the advantage of converting waste carbon-dioxide, a greenhouse gas to environmental friendly plastics using the energy of sunlight. Materials and Methods Sample collection Samples were collected from diferent freshwater habitats of Tamilnadu which includes Muttukaddu lake, Elagiri lake, Tuticorin; Panderavellai; Alandur and WCC pond water. Isolation and purifcation Te algal species were isolated and purifed from diferent sources using basic microbial techniques primarily with serial dilution and followed by spread plating and quadrant streaking on BBM (Bold’s Basal Medium) agar plates. Morphological identifcation was performed through microscopic observations. Culture conditions Te purifed axenic cultures were grown in 250 ml Erlenmeyer fasks containing 100 ml of BBM. Experimental cultures were incubated at 25+2°C, 14/10 light/dark cycles with illumination of 3000 lux under cool white fuorescent lamps. Every day the cultures were mildly shaken by hand for 10 minutes. Screening for the production of PHB using sudan black staining technique Te isolates were stained with Sudan Black stain. Te samples were stained for 10 mins with Sudan Black Solution, rinsed with water and counter stained with 0.5% safranin for 5 mins. Te slide was observed under the microscope at 1000 x magnifcation. Nile blue staining technique Heat fxed cells were treated with 1% Nile blue for 10 minutes and was observed at an excitation wavelength of 460 nm. Analytical procedure Te algal cultures were centrifuged at 10,000 rpm for 5 minutes to obtain cell pellet. Te pellet was used to estimate the wet cell weight (WCW in g/ml) [3] and dry cell weight (DCW in g/ml).   % =  ℎ    /  / × 100 Optimization of media Te following media was prepared and the isolated six Chlorella vulgaris PB (1-6) obtained from diferent sources were inoculated and incubated at 14/10 light/dark cycle with illumination of 3000 lux under J o u r n a l o f B i o r e m e d i a ti o n & B i o d e g r a d a t i o n ISSN: 2155-6199 Journal of Bioremediation & Biodegradation Robert and Iyer, J Bioremediat Biodegrad 2018, 9:2 DOI: 10.4172/2155-6199.1000433 Research Article Open Access J Bioremediat Biodegrad, an open access journal ISSN: 2155-6199 Volume 9 • Issue 2 • 1000433