Optimization of Vertical Fixed-Bed Pyrolysis for Enhanced Biochar Production from Diverse Agricultural Residues.

This study examines the pyrolysis of agricultural residues, namely, coconut shells, rice husks, and cattle manure, in a vertical fixed-bed reactor at varying temperatures from 300 to 800 degrees Celsius for biochar production. The research aimed to evaluate the potential of biochar as biofuels, adsorbents, and soil amendments. Proximate, ultimate, and elemental analyses were conducted to determine their composition and caloric values. Several analytical techniques were used in the physical and chemical characterization of the biochar (SEM, FTIR, BET). The results indicated that the highest SBET values were achieved under different conditions for each biochar: 89.58 m2/g for BC-CS-700, 202.39 m2/g for BC-RH-600, and 42.45 m2/g for BC-CD-800. Additionally, all three biochars exhibited the highest caloric values at 600 °C. The results showed that 600 °C is the general optimal temperature to produce biochar from an assortment of biomass materials, considering their use for a variety of purposes. BC-CS-800 had the highest elemental carbon content at 93%, accompanied by a relative decrease in oxygen content. The van Krevelen diagram of biochar products shows that biochars derived from coconut shells and rice husks are suitable for use as fuels. Furthermore, FTIR analysis revealed the presence of oxygen-containing functional groups on the biochar surface, enhancing their pollutant adsorption capabilities. This study provides valuable insights into the scalable and environmentally sustainable production of biochar, emphasizing its role in improving soil quality, increasing energy density, and supporting sustainable agricultural practices.

Pathways of soil organic carbon accumulation are related to microbial life history strategies in fertilized agroecosystems.

Although the formation, turnover, and accumulation of soil organic carbon (SOC) are driven by different fertilizer inputs and their subsequent microbial-mediated transformation, the relationship between changes in plant-derived and microbial-derived components and soil microbial life history strategies under different fertilization regimes has not been well explored. In this study, the changes in microbial necromass carbon (MNC), lignin phenols, and glomalin-related soil protein (GRSP), as well as soil microbial life history strategy were determined in a 16-year field experiment in response to different fertilization regimes, including a no-fertilizer control (C), conventional chemical NPK fertilization (NPK), and partial substitutions of the NPK in chemical fertilizers with a low (30 %) or high (60 %) level of straw (0.3S and 0.6S) or cattle manure (0.3M and 0.6M). The results showed that total lignin phenol content and its contribution to SOC were significantly increased by 88.7 % and 74.2 %, respectively, in high-level straw substitution treatment as compared to chemical fertilization. Both high-level straw and cattle manure substitution increased MNC and total GRSP contents, but did not alter their contributions to SOC compared to chemical fertilization. In fertilized treatments, the high-level cattle manure substitution had the lowest and highest bacterial and fungal K/r ratio, respectively. Bacterial K/r ratio was an important factor in predicting bacterial necromass carbon content and there was a significant negative correlation between them. The ratio of ectomycorrhizal to saprotrophic fungi and fungal diversity were important factors for predicting lignin phenol and GRSP contents, respectively. In addition, the SEMs modeling indicated that straw substitution directly affected lignin phenol and MNC accumulation, whereas cattle manure substitution indirectly affected MNC accumulation by affecting microbial life history strategies. In conclusions, agricultural residues inputs support the formation of a multiple carbon pool of SOC compared to chemical fertilization; and microbial life history strategy is an important driver of SOC formation and affects SOC accumulation and stability in agroecosystems.

Experimental investigation on utilization of Sesbania grandiflora residues through thermochemical conversion process for the production of value added chemicals and biofuels.

All the countries in the world are now searching for renewable, environmentally friendly alternative fuels due to the shortage and environmental problems related with the usage of conventional fuels. The cultivation of cereal and noncereal crops through agricultural activities produces waste biomasses, which are being evaluated as renewable and viable fossil fuel substitutes. The thermochemical properties and thermal degradation behavior of Sesbania grandiflora residues were investigated for this work. A fluidized bed reactor was used for fast pyrolysis in order to produce pyrolysis oil, char and gas. Investigations were done to analyze the effect of operating parameters such as temperature (350-550 °C), particle size (0.5-2.0 mm), sweeping gas flow rate (1.5-2.25 m3/h). The maximum of pyrolysis oil (44.7 wt%), was obtained at 425 °C for 1.5 mm particle size at the sweep gas flow rate of 2.0 m3/h. Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry methods were used to examine the composition of the pyrolysis oil. The pyrolysis oil is rich with aliphatic, aromatic, phenolic, and some acidic chemicals. The physical characteristics of pyrolysis oil showed higher heating value of 19.76 MJ/kg. The char and gaseous components were also analyzed to find its suitability as a fuel.

Hydrogen and methane production through two stage anaerobic digestion of straw residues.

Anaerobic digestion of agricultural waste can contribute to the European renewable energy needs. The 71% of the 20,000 anaerobic digestion plants in operation already uses these agro-waste as feedstock; part of these plants can be converted into two stage processes to produce hydrogen and methane in the same plant. Biomethane enriched in hydrogen can replace natural gas in grids while contributing to the sector decarbonisation. Straw is the most abundant agricultural residue (156 Mt/y) and its conventional final fate is uncontrolled soil disposal, landfilling, incineration or, in the best cases, composting. The present research work focuses on the fermentation of spent mushroom bed, an agricultural lignocellulosic byproduct, composed mainly from wheat straw. The substrate has been characterized and semi-continuous tests were performed evaluating the effect of the hydraulic retention time on hydrogen and volatile fatty acids production. It was found that all the tests confirmed the feasibility of the process even on this lignocellulosic substrate, and also, it was identified HRT 4.0 d as the best option to optimize the productivity of volatile fatty acids (17.09 gCODVFAs/(KgVS*d)), and HRT 6.0 d for hydrogen (7.98 LH2/(KgVS*d)). The fermentation effluent was used in biomethanation potential tests to evaluate how this process affects a subsequent digestion phase, reporting an increase in the energetical feedstock exploitation up to 30%.

Investigation on thermochemical co-gasification of rice husk and groundnut shell in open core gasifier for the generation of producer gas: An optimization by central composite design.

Several energy-related strategies and scenarios have been suggested to address concerns about rising global temperatures. In addition to using renewable energy, the improvement in energy efficiency of conventional systems is also in focus. Policies are already in place in many countries, including India, to address the energy needs of rural and small-scale enterprises by gasifying locally available, diverse agricultural leftovers. Although rice husk and groundnut shell are two commonly used agricultural leftovers in the southern part of India, their appropriate blending must be studied to improve their conversion efficiency in co-gasification. Therefore, the primary objective of this research is to construct a statistical model utilizing response surface methodology (RSM) to analyze the thermochemical co-gasification of the aforementioned biomass materials. Since RSM can predict optimum performance with limited experimental data, this could contribute to the identification of the performance and operating parameters of an open-core gasifier. The model predicts that the mixture containing 20% rice husk and working at an ER of 0.25 and a reduction zone inlet temperature of 879.9 °C will be CO-23.53%, H2-13.97%, and CH4-3.56%. In addition, the lower heating value and gas yield can be as high as 6.17 MJ/Nm3 and 2.369 m3/kg, respectively. This outcome can contribute to the effective utilization of biomass for energy supply in rural areas. However, the economic parameters must be analyzed to implement the same in any region.

Emission characteristics of carbonyl compounds from open burning of typical subtropical biomass in South China.

Open biomass burning (OBB) is one of the largest primary emission sources for atmospheric carbonyl compounds, key precursors for ozone and secondary organic aerosol pollution. To clarify the carbonyl emissions, the comprehensive characteristics of C1-C10 carbonyl compounds from open burning of seven typical subtropical biomass in China were investigated in this study, which included subtropical plants and agricultural residues. Total 27 carbonyl compounds were detected. The total EFs were 2824 mg kg-1 with 95% confidence interval (CI) [2418, 3322] for burning subtropical plants and 4080 mg kg-1 with 95% CI [3446, 4724] for burning agriculture residues, respectively. The EFs were 2-3 orders of magnitude larger than previous values in China. Aliphatic aldehydes were the largest group of carbonyl groups, with acetaldehyde, as the most abundant carbonyl species (about 30% contribution). Formaldehyde, acetone, acrolein, glyoxal, methylglyoxal, butanone, isovaleraldehyde, and m-tolualdehyde were also found to be abundant and varying with the types of biomass burnt. Formaldehyde emission ratios to acetonitrile and CO were lower than those in previous studies both for burning plants and agricultural residues. There were significant variabilities in the emission ratios and factors among different types of OBBs. Strong positive correlations were found between carbonyl emissions and CO emissions and water content in biomass; furthermore, total carbonyl concentrations measured in the flaming stage were higher than those in the smoldering one. This study provides important fundamental measurement data on carbonyl emissions from burning typical subtropical plants and agricultural residues, which will help improve the quality of emission inventories and better understand the potential impacts of OBB on regional air quality in southern China.

Effect of Enzymatic Biotransformation on the Hypotensive Potential of Red Grape Pomace Extract.

Hypertension is a widespread health risk, affecting over a billion people and causing 9 million deaths per year. The Renin-Angiotensin-Aldosterone System (RAAS) is a primary target for hypertension treatment, and it is primarily treated through drugs that inhibit the Angiotensin I-Converting Enzyme (ACE). In addition to pharmacological treatment, various plants are recommended in traditional medicine for blood pressure regulation. This study aimed to produce high-phenolic-content extracts with and without enzymatic assistance from red grape pomace and evaluate their antioxidant capacity and ACE inhibitory potential. The total phenolic content (TPC) was measured, and phenolic identification was performed using HPLC analysis. In addition, the antioxidant capacity and anti-hypertensive potential were determined via in vitro assays. There was no statistical difference in the TPC antioxidant capacity between the extraction methods. Otherwise, when considering the extraction yield, the enzymatic process recovered around 70% more phenolic compounds from the pomace, and the phenolic profile was changed. Enzymatic assistance also significantly increased the ACE inhibitory potential in the grape pomace extract. This study demonstrates the viability of upcycling grape pomace to obtain bioactive compounds and to reduce their environmental impact, and highlights the influence of the enzymatic extraction on the hypotensive potential of the extract.

Garbage to Gains: The role of biochar in sustainable soil quality improvement, arsenic remediation, and crop yield enhancement.

Threats of soil quality deterioration and metal pollution have inflicted several parts of the world, apart from the need for surplus crop production. The investigation used biochar prepared from waste biomasses such as wheat and rice straw, kitchen waste, leaf litter, Lantana camara, orange peel, and walnut shell to improve soil quality, reduce As pollution, and enhance plant growth. Biochars were amended at doses of 0%, 2.5%, 5%, and 7.5% and conditioned for 3 months. At a 7.5% dose, the maximum improvements in cation exchange capacity (a 62% increase), anion exchange capacity (a two-fold increase), bulk density (a 31% decrease), porosity (a 32% increase), water holding capacity (an 86% increase), soil respiration (a 32% increase), total carbon (a two-and-a-half-fold increase), total nitrogen (an eleven-fold increase), total phosphorus (3 times rise), total potassium (a two-and-a-half-fold increase), mobile As (a 38% decrease), leachable As (a 53% decrease), and bio-available As (a 56% decrease) were observed. Further, pot experiments revealed augmented biomass growth (61% and 177%), increased length (71% and 209%), and decreased As accumulation (56% and 55%) in the above-ground parts of Bengal gram and coriander plants, respectively. Therefore, the application of biochar was found to enhance the physico-chemical properties of soil, reduce As contamination levels, and improve crop growth. The study recommends using waste biomasses to prepare eco-friendly biochars, which could contribute to advancing sustainable agriculture and the circular economy.

Evaluation of commercial moving bed media and sugarcane bagasse as packing material in biotrickling filter for hydrogen sulfide removal.

This study compared two biotrickling filter packing materials for hydrogen sulfide removal. Inlet H2S concentrations and empty-bed retention time were tested on the two biotrickling filters. First reactor (BT1) had immobilized sulfur-oxidizing bacteria on commercial moving-bed media, whereas second reactor (BT2) had sulfur-oxidizing bacteria on sugarcane bagasse. The study found that BT1 performed best at 120 s empty-bed retention time, 422.39 g/m3·h hydrogen sulfide loading rate, resulted in 416 g/m3·h hydrogen sulfide elimination capacity. In contrast, BT2 performed best at 180 s empty-bed retention time, 278.77 g/m3·h hydrogen sulfide loading rate, and 273 g/m3·h elimination capacity was achieved. High-throughput sequencing showed Acidithobacillus spp. dominated the sulfur-oxidizing bacteria consortium. Sugarcane bagasse may receive less hydrogen sulfide loading than moving bed medium under optimal conditions, but its low cost and reasonable removal capacity of hydrogen sulfide -containing industrial gases in a biotrickling filter system make it an excellent alternative packing material.

Data on exopolysaccharides produced by Bacillus spp. from cassava pulp with antioxidant and antimicrobial properties.

This data evaluated the capacity of Bacillus spp. isolated from Thai milk kefir to produce exopolysaccharide (EPS) on cassava pulp and tested its antioxidant and antibacterial properties. Thailand's starch industry generates million tons of cassava pulp, which is underutilized or bio-transformed into higher-value bioproducts. Antioxidant and antibacterial bacterial exopolysaccharides are beneficial in the food, feed, pharmaceutical, and cosmetic industries. Moisture, ash, fat, protein, fiber, starch, sugar, neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL) were analyzed from cassava pulp as an EPS substrate. After 3 days of bacterial fermentation, EPS generation, culture pH, reducing sugar amount, and bacterial count were recorded. Antioxidant activities and bioactive content including hydroxyl radical scavenging activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP), total phenolic and flavonoid content (TPC and TFC), and antimicrobial activity against two Nile tilapia pathogens (Streptococcus agalactiae and Staphylococcus aureus) from different Bacillus species were evaluated. Proximate analysis, dinitrosalicylic acid assay, pH value record, bacterial count using spread plate method, antioxidant activity and bioactive content assays via spectrophotometry, and agar disk diffusion were the main approaches. This study used microbial cell factories to convert agro-biowaste, such as cassava pulp, into EPS bioproducts which accords with a bio-circular green economy model.

Exploring increased hydraulic retention time as a cost-efficient way of valorizing residual biogas potential.

Effective substrate utilization with low residual methane yield in the digestate is crucial for the economy and sustainability of biogas plants. The composition and residual methane potential of 29 digestate samples from plants operating at hydraulic retention times of 13-130 days were determined to evaluate the economic viability of extended digestion. Considerable contents of fermentable fractions, such as cellulose (8-23%), hemicellulose (1-18%), and protein (13-22%), were present in the digestate dry matter. The ultimate residual methane yields varied between 55 and 236 ml/g of volatile solids and correlated negatively with the logarithm of the hydraulic retention time (r = -0.64, p < 0.05). Economic analysis showed that extending the retention time in 20 days would be viable for 18 systems if methane were sold for 1.00 €/m3, with gains up to 40 €/year/m3 of newly installed reactor capacity. The results show the importance of operating at sufficient hydraulic retention time.

Cellulose nanocrystals from agricultural residues (Eichhornia crassipes): Extraction and characterization.

Extraction of cellulose nanocrystals (CNCs) from agro-residues has received much attention, not only for their unique properties supporting a wide range of potential applications, but also their limited risk to global climate change. This research was conducted to assess Nile roses (Eichhornia crassipes) fibers as a natural biomass to extract CNCs through an acid hydrolysis approach. Nile roses fibers (NRFs) were initially subjected to alkaline (pulping) and bleaching pretreatments. Microcrystalline cellulose (MCC) was used as control in comparison to Nile rose based samples. All samples underwent acid hydrolysis process at a mild temperature (45 °C). The impact of extraction durations ranging from 5 to 30 min on the morphology structure and crystallinity index of the prepared CNCs was investigated. The prepared CNCs were subjected to various characterization techniques, namely: X-ray diffraction (XRD), FT-IR analysis, Transmission electron microscopy (TEM), and X-ray Photoelectron spectroscopy (XPS). The outcomes obtained by XRD showed that the crystallinity index increased as the duration of acid hydrolysis was prolonged up to 10 min, and then decreased, indicating optimal conditions for the dissolution of amorphous zones of cellulose before eroding the crystallized domains. These data were confirmed by FT-IR spectroscopy. However, a minor effect of hydrolysis duration on the degree of crystallinity was noticed for MCC based samples. TEM images illustrated that a spherical morphology of CNCs was formed as a result of 30 min acid hydrolysis, highlighting the optimal 20 min acid hydrolysis to obtain a fibrillar structure. The XPS study demonstrated that the main constituents of extracted CNCs were carbon and oxygen.

Upcycling of PHA-producing bacteria for biostimulants production and polyhydroxyalkanoates recovery.

The use of petroleum-based plastic has led to its accumulation in the environment, with negative impacts on the ecosystem and the biota. Polyhydroxyalkanoates (PHAs), biobased and biodegradable plastics produced by microbes, have many commercial applications, however their high production cost means they cannot yet compete with traditional plastics. At the same time, the problem of the growing human population implies that improved crop production is needed to avoid malnutrition. Biostimulants enhance plant growth and have the potential to improve agricultural yields; they can be obtained from biological feedstock, including microbes. Therefore, there is scope for coupling the production of PHAs with that of biostimulants, making the process more cost-efficient and minimizing by-products generation. In this work, low-value agro-zoological residues were processed to obtain PHA-storing bacteria via acidogenic fermentation; PHAs destined for the bioplastic market were extracted, and the protein-rich by-products were turned into protein hydrolysates using different treatment methods, assessing their biostimulant effects in growth trials with tomato and cucumber plants. The results indicate that the best hydrolysis treatment, realizing the highest amount of organic nitrogen (6.8 gN-org/L) while achieving the best PHA recovery (63.2 % gPHA/gTS), is obtained with strong acids. All the protein hydrolysates were effective in improving either roots or leaf development, with various results, depending on the species and the growth method. The acid hydrolysate was the most effective treatment to enhance the development of shoots (21 % increase compared to the control) and roots (16 % increase for the dry weight and 17 % for main root length) of hydroponically-grown cucumber plants, while pot-grown tomatoes, biostimulated via foliar spray, developed bigger shoots (up to 41 %) with the hydrolysate obtained from the alkaline treatment. These preliminary results indicate that simultaneous production of PHAs and biostimulants is feasible, and that commercialization could be achievable given the expected reduction in production costs.

High-level production of Aspergillus niger prolyl endopeptidase from agricultural residue and its application in beer brewing.

BACKGROUND: Prolyl endopeptidase from Aspergillus niger (AN-PEP) is a prominent serine proteinase with various potential applications in the food and pharmaceutical industries. However, the availability of efficient and low-cost AN-PEP remains a challenge owing to its low yield and high fermentation cost. RESULTS: Here, AN-PEP was recombinantly expressed in Trichoderma reesei (rAN-PEP) under the control of the cbh1 promoter and its secretion signal. After 4 days of shaking flask cultivation with the model cellulose Avicel PH101 as the sole carbon source, the extracellular prolyl endopeptidase activity reached up to 16.148 U/mL, which is the highest titer reported to date and the secretion of the enzyme is faster in T. reesei than in other eukaryotic expression systems including A. niger and Komagataella phaffii. Most importantly, when cultivated on the low-cost agricultural residue corn cob, the recombinant strain was found to secret a remarkable amount of rAN-PEP (37.125 U/mL) that is twice the activity under the pure cellulose condition. Furthermore, treatment with rAN-PEP during beer brewing lowered the content of gluten below the ELISA kit detection limit (< 10 mg/kg) and thereby, reduced turbidity, which would be beneficial for improving the non-biological stability of beer. CONCLUSION: Our research provides a promising approach for industrial production of AN-PEP and other enzymes (proteins) from renewable lignocellulosic biomass, which provides a new idea with relevant researchers for the utilization of agricultural residues.

The effect of biochar prepared at different pyrolysis temperatures on microbially driven conversion and retention of nitrogen during composting.

Aerobic composting is one of the most economical ways to produce organic fertilizer from agricultural wastes. In this research, we independently developed a simple composting simulation reactor. The effects of biochar pyrolysised at different pyrolysis temperatures (B1-450 °C; B2-550 °C; and B3-650 °C) on nitrogen conversion (Total nitrogen (TN), ammonium nitrogen (NH4 +-N), nitrate nitrogen (NO3 --N), cumulative amount of ammonia (CEA) and nitrous oxide (CEN) emission, nitrogen loss rate (NLR), etc.) and functional microbial community (cbbL, cbbM and nifH) structure in the composting system were studied. Results showed that the addition of biochar significantly improved the efficiency of composting, increased the NO3 --N concentration and reduced the NLR (%) in the composting system (B3 (31.4 ± 2.73)

Bioethanol production from agricultural residues as lignocellulosic biomass feedstock's waste valorization approach: A comprehensive review.

Bioenergy is becoming very popular, drawing attention as a renewable energy source that may assist in managing growing energy costs, besides possibly affording revenue to underprivileged farmers and rural populations worldwide. Bioethanol made from agricultural residual-biomass provides irreplaceable environmental, socioeconomic, and strategic benefits and can be considered as a safe and cleaner liquid fuel alternative to traditional fossil fuels. There is a significant advancement made at the bench scale towards fuel ethanol production from agricultural lignocellulosic materials (ALCM). These process technologies include pretreatment of ALCM biomass employment of cellulolytic enzymes for depolymerizing carbohydrate polymers into fermentable sugars to effectively achieve it by applying healthy fermentative microbes for bioethanol generation. Amongst all the available process methods, weak acid hydrolysis followed by enzymatic hydrolysis process technique. Recovering higher proficient celluloses is more attractive in terms of economic benefits and long-term environmental effects. Besides, the state of ALCM biomass based bioethanol production methods is discussed in detail, which could make it easier for the scientific and industrial communities to utilize agricultural leftovers properly.

Agricultural Residues as Raw Materials for Pulp and Paper Production: Overview and Applications on Membrane Fabrication.

The need for pulp and paper has risen significantly due to exponential population growth, industrialization, and urbanization. Most paper manufacturing industries use wood fibers to meet pulp and paper requirements. The shortage of fibrous wood resources and increased deforestation are linked to the excessive dependence on wood for pulp and paper production. Therefore, non-wood substitutes, including corn stalks, sugarcane bagasse, wheat, and rice straw, cotton stalks, and others, may greatly alleviate the shortage of raw materials used to make pulp and paper. Non-woody raw materials can be pulped easily using soda/soda-AQ (anthraquinone), organosolv, and bio-pulping. The use of agricultural residues can also play a pivotal role in the development of polymeric membranes separating different molecular weight cut-off molecules from a variety of feedstocks in industries. These membranes range in applications from water purification to medicinal uses. Considering that some farmers still burn agricultural residues on the fields, resulting in significant air pollution and health issues, the use of agricultural residues in paper manufacturing can eventually help these producers to get better financial outcomes from the grown crop. This paper reviews the current trends in the technological pitch of pulp and paper production from agricultural residues using different pulping methods, with an insight into the application of membranes developed from lignocellulosic materials.

Formation and application of edible oleogels prepared by dispersing soy fiber particles in oil phase.

Oleogels containing less saturated and trans-fats were considered as an ideal option to replace the solid fats in foods. In this research, oleogel was fabricated by dispersing soy fiber particles (SFP) in soy oil, and further it was used in bread preparation. Effect of the particle size, particle content and the second fluid content on the formation of oleogels were evaluated, based on the appearance and rheological properties. Results showed that the suspension of SFP in soy oil (24%, w/w) could be transformed into gel-like state, upon the addition of the second fluid. The SFP based networks were dominated by the capillary force which was originated from the second fluid. The rheological properties and yield stress of the oleogels could be modulated by particle size and particle content of SFP in oil phase, as well as the second fluid content in the system. When the oleogels were applicated in bread preparation, a layered structure could be formed in the bread, indicating the possibility of replacing the solid fats in bakery products by our oleogels. Our results offered a feasibility approach for oil structuring with natural raw materials, and developed a new approach to replace the solid fats in foods.

Cost effective media optimization for PHB production by Bacillus badius MTCC 13004 using the statistical approach.

Polyhydroxybutyrate (PHB) has significant potential for replacing non-biodegradable traditional plastic, which is responsible for several global environmental issues. The main problem with switching to bio-based alternatives for petrochemical plastics is the large price gap on the market. To overcome this problem, the present research was focused on the utilization of inexpensive substrates i.e. agricultural residues for cost-effective PHB production by endospore-forming bacteria Bacillus badius MTCC 13004. For efficient PHB production, Box-Behnken Design (BBD) was selected for media optimization and to observe the interactive effects of four variables i.e. pH, Na acetate, Banana peel, and mustard cake. PHB yield of 2.11 g/L was attained under optimized conditions compared to non-optimized conditions (0.72 g/L). FTIR spectra analysis of PHB extracted from Bacillus badius was found to be similar to commercial PHB. NMR data was also matched with the chemical shift signals CH, CH2, and CH3 of PHB. The melting temperature (Tm) and glass transition temperature (Tg) of PHB from Bacillus badius was found to be 165.14 and 2.68 °C, respectively. Further, PCR protocol was also designed to amplify key enzymes of the PHB synthesis pathway i.e. PHB synthase (phb C gene).