Efficient biodegradation of Polyethylene terephthalate (PET) plastic by Gordonia sp. CN2K isolated from plastic contaminated environment.

Since we rely entirely on plastics or their products in our daily lives, plastics are the invention of the hour. Polyester plastics, such as Polyethylene Terephthalate (PET), are among the most often used types of plastics. PET plastics have a high ratio of aromatic components, which makes them very resistant to microbial attack and highly persistent. As a result, massive amounts of plastic trash accumulate in the environment, where they eventually transform into microplastic (<5 mm). Rather than macroplastics, microplastics are starting to pose a serious hazard to the environment. It is imperative that these polymer microplastics be broken down. Through the use of enrichment culture, the PET microplastic-degrading bacterium was isolated from solid waste management yards. Bacterial strain was identified as Gordonia sp. CN2K by 16 S rDNA sequence analysis and biochemical characterization. It is able to use polyethylene terephthalate as its only energy and carbon source. In 45 days, 40.43 % of the PET microplastic was degraded. By using mass spectral analysis and HPLC to characterize the metabolites produced during PET breakdown, the degradation of PET is verified. The metabolites identified in the spent medium included dimer compound, bis (2-hydroxyethyl) terephthalate (BHET), mono (2-hydroxyethyl) terephthalate (MHET), and terephthalate. Furthermore, the PET sheet exposed to the culture showed considerable surface alterations in the scanning electron microscope images. This illustrates how new the current work is.

Biodegradation of fluoranthene by Paenibacillus sp. strain PRNK-6: a pathway for complete mineralization.

A high-efficiency fluoranthene-degrading bacterium Paenibacillus sp. PRNK-6 was isolated from PAH-contaminated soil. The strain degrades 96% (240 mg l-1) of fluoranthene in 48 h. Various metabolic intermediates of fluoranthene catabolism were identified by gas chromatography (GC) and gas chromatography-high resolution mass spectrometry (GC-HRMS). Metabolite characterization, metabolite-feeding experiments, and appropriate enzyme activities in the cell-free extracts suggest the existence of a bifurcated pathway down the phthalic acid for complete mineralization of fluoranthene in PRNK-6. In this strain, fluoranthene catabolism begins by the attack on the fused aromatic ring portion of fluoranthene. Two terminal aromatic metabolites protocatechuate and catechol undergo ring cleavage by protocatechuate 3,4-dioxygenase and catechol 1,2-dioxygenase, respectively, and enter the central metabolism.

Enhanced utilization of fluorene by Paenibacillus sp. PRNK-6: Effect of rhamnolipid biosurfactant and synthetic surfactants.

The present investigation was to study the effect of different non-ionic surfactants (Tween-80, Tween-60, Tween-40, Tween-20, Triton X-100) and a rhamnolipid biosurfactant on the degradation of fluorene by Paenibacillus sp. PRNK-6. An enhancement in the growth, as well as fluorene utilization by this strain were observed in the presence of biosurfactant and non-ionic surfactants except Tween-20 and Triton X-100. Triton X-100 and Tween-20 were toxic to this bacterium. The strain PRNK-6 utilized 75% of fluorene (280mg/L) in 24h in an unamended condition. On the other hand, the complete utilization of higher concentration fluorene (320mg/L) by this strain was noticed when the medium was amended with Tween-80 (1.5% v/v) within 24h of incubation. Whereas, 90.6%, 96.5% and 96.7% of fluorene (280mg/L) was utilized when amended with Tween-60 (3.5% v/v), Tween-40 (3% v/v) and biosurfactant (25mg/L) respectively. Biosurfactant promoted the fluorene degradation potential of PRNK-6 as 96.2% of 320mg/L fluorene was degraded within 24h. Further, the added tween series surfactants and a biosurfactant have increased the cell surface hydrophobicity of the PRNK-6. Thus correlating with the enhanced degradation of the fluorene.

A plate method for screening of bacteria capable of degrading aliphatic nitriles.

A novel indicator plate method was developed for screening of aliphatic-nitrile-degrading bacteria. Isolated bacteria were tested for utilization of acetonitrile as sole source of carbon and nitrogen with the release of ammonia. The released ammonia causes increase of the pH of the medium. Phenol red indicator is used for detection of ammonia based on colour change of the indicator dye from red to pink. The liberation of ammonia from aliphatic-nitrile-utilizing bacteria is also studied in plates containing other indicators such as bromothymol blue and phenolphthalein. The usefulness of the indicator plate is demonstrated for bacteria that degrade certain aliphatic nitriles. Bacteria degrading nitriles as a nitrogen source can also be isolated with a medium containing additional carbon source. This plate method would be useful in isolation and screening of bacteria for degradation of aliphatic nitriles and also for production of nitrile-hydrolyzing enzymes.

Metabolism of acenaphthylene via 1,2-dihydroxynaphthalene and catechol by Stenotrophomonas sp. RMSK.

Stenotrophomonas sp. RMSK capable of degrading acenaphthylene as a sole source of carbon and energy was isolated from coal sample. Metabolites produced were analyzed and characterized by TLC, HPLC and mass spectrometry. Identification of naphthalene-1,8-dicarboxylic acid, 1-naphthoic acid, 1,2-dihydroxynaphthalene, salicylate and detection of key enzymes namely 1,2-dihydroxynaphthalene dioxygenase, salicylaldehyde dehydrogenase and catechol-1,2-dioxygenase in the cell free extract suggest that acenaphthylene metabolized via 1,2-dihydroxynaphthalene, salicylate and catechol. The terminal metabolite, catechol was then metabolized by catechol-1,2-dioxygenase to cis,cis-muconic acid, ultimately forming TCA cycle intermediates. Based on these studies, the proposed metabolic pathway in strain RMSK is, acenaphthylene --> naphthalene-1,8-dicarboxylic acid --> 1-naphthoic acid --> 1,2-dihydroxynaphthalene --> salicylic acid --> catechol --> cis,cis-muconic acid.

Degradation of cinnamic acid by a newly isolated bacterium Stenotrophomonas sp. TRMK2.

A bacterium Stenotrophomonas sp. TRMK2 capable of utilizing cinnamic acid was isolated from agro-industrial waste by enrichment culture technique. This strain completely utilizes 5 mM cinnamic acid within 18 h of incubation. The different metabolites formed during the degradation of cinnamic acid were characterized by GC-HRMS. The involvement of various enzymes, namely cinnamate reductase, 3-phenylpropionic acid hydroxylase, p-hydroxybenzoic acid hydroxylase and protocatechuate 3,4-dioxygenase in cinnamic acid degradation was demonstrated. A catabolic pathway for cinnamic acid in Stenotrophomonas sp. TRMK2 is as follows: Cinnamic acid; 3-Phenylpropionic acid; 3-(4-Hydroxyphenyl) propionic acid; 4-Hydroxy benzoic acid and Protocatechuic acid. Further, this strain is capable of utilizing various phenolic compounds.

A comparative study of utilization of single and mixed phenolic compounds by individual and mixed culture.

Three bacterial strains; Pseudomonas sp. TRMK1, Stenotrophomonas sp. TRMK2 and Xanthomonas sp. TRMK3 were isolated from agro-industrial waste by enrichment culture technique that are capable of utilizing phenolic acids as sole source of carbon and energy. These strains were found to utilize p-coumaric, ferulic and caffeic acid. The individual strains utilized 5 mM of mixed phenolic acids within 20 h of incubation. The bacterial consortium composing these strains was prepared and studied the efficient degradation of phenolic compounds. The bacterial consortium showed the enhanced utilization of 30 mM individual and 25 mM mixed phenolic acids within 32 and 40 h of incubation, respectively. The degradation efficiency of these strains in all the above experiments was above 90%. The prepared bacterial consortium serves as a suitable method for the in situ application of sites contaminated with wide range of phenolic compounds.

Utilization of Phenylpropanoids by Newly Isolated Bacterium Pseudomonas sp. TRMK1.

A bacterium Pseudomonas sp. TRMK1 capable of utilizing various phenylpropanoids was isolated from agro-industrial waste by enrichment culture technique. It is gram-negative, motile, aerobic, and able to utilize three different phenolic acids such as p-coumaric, ferulic, and caffeic acids at concentrations of 5, 10, and 15 mM in 18 h of incubation. The residual concentration of phenolic acids was analyzed by HPLC. The catabolic pathway of p-coumaric, ferulic, and caffeic acids is suggested based on the characterization of metabolic intermediates by GC, GC-HRMS, and different enzymatic assays. Further, Pseudomonas sp. TRMK1 utilizes a wide range of mixture of phenolic acids present in the synthetic effluent.

Biodegradation of butyronitrile and demonstration of its mineralization by Rhodococcus sp. MTB5.

A nitrile utilizing bacterium Rhodococcus sp. MTB5 was previously isolated in our laboratory by the enrichment culture technique. It is able to utilize butyronitrile as sole carbon, nitrogen, and energy source. Maximum butyronitrile degrading property of this strain has been investigated. Results reveal that 100, 98, and 88 % degradation was achieved for 2, 2.5, and 3 % butyronitrile, respectively. The strain is capable of growing in as high as 5 % butyronitrile concentration. A two-step pathway involving nitrile hydratase (NHase) and amidase was observed for the biodegradation of butyronitrile. Complete degradation (mineralization) of butyronitrile with the help of metabolite feeding experiment was reported. The significance of this investigation was the capability of the strain to completely degrade and its ability to grow on higher concentrations of butyronitrile. These potential features make it a suitable candidate for practical field application for effective in situ bioremediation of butyronitrile contaminated sites.

A catabolic pathway for the degradation of chrysene by Pseudoxanthomonas sp. PNK-04.

The chrysene-degrading bacterium Pseudoxanthomonas sp. PNK-04 was isolated from a coal sample. Three novel metabolites, hydroxyphenanthroic acid, 1-hydroxy-2-naphthoic acid and salicylic acid, were identified by TLC, HPLC and MS. Key enzyme activities, namely 1-hydroxy-2-naphthoate hydroxylase, 1,2-dihydroxynaphthalene dioxygenase, salicylaldehyde dehydrogenase and catechol-1,2-dioxygenase, were noted in the cell-free extract. These results suggest that chrysene is catabolized via hydroxyphenanthroic acid, 1-hydroxy-2-naphthoic acid, salicylic acid and catechol. The terminal aromatic metabolite, catechol, is then catabolized by catechol-1,2-dioxygenase to cis,cis-muconic acid, ultimately forming TCA cycle intermediates. Based on these studies, the proposed catabolic pathway for chrysene degradation by strain PNK-04 is chrysene → hydroxyphenanthroic acid → 1-hydroxy-2-naphthoic acid → 1,2-dihydroxynaphthalene → salicylic acid → catechol →cis,cis-muconic acid.