Micropollutants (ciprofloxacin and norfloxacin) remediation from wastewater through laccase derived from spent mushroom waste: Fate, toxicity, and degradation.

Fluoroquinolone antibiotics frequently found in environmental matrices (wastewater treatment plants, hospital wastewater, industrial wastewater and surface wastewater) causes potential threat to the environment. Enzymatic treatment for degradation of antibiotics from environmental matrices is a green and sustainable approach. Focusing on this, this study aimed to degrade two frequently found fluroquinolone emergent pollutants, ciprofloxacin and norfloxacin from wastewater. The trinuclear cluster of copper ions present in laccase has the ability to effectively remove organic micropollutants (OMPs). The uniqueness of this study is that it utilizes laccase enzyme extracted from spent mushroom waste (SMW) of P. florida for degradation of ciprofloxacin and norfloxacin and to achieve highest degradation efficiency various parameters were tweaked such as pH (3-6), temperature (30 °C and 50 °C), and ABTS (0.05, 0.6, and 1 mM) concentration. The results showed that the most effective degradation of ciprofloxacin (86.12-75.94%) and norfloxacin (83.27-65.94%) was achieved in 3 h at pH 4.5, temperature 30 °C, and 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 0.05 mM concentration. Nevertheless, achieving degradation at 50 °C for both antibiotics, indicates thermostability nature of laccase (P. florida). Further, the fate of transformed products obtained from laccase mediated degradation was confirmed by liquid chromatography (LC-MS). Both the antibiotics undergo decarboxylation, depiperylyzation, dealkylation and defluorination as a result of laccase-mediated bond breakage. Anti-microbial activity of the biodegraded products was monitored by residual anti-bacterial toxicity test (E. coli and Staphylococcus aureus). The biodegraded products were found to be non-toxic and resulted in the growth of E. coli and Staphylococcus aureus, as determined by the agar-diffusion method. Moreover, the storage stability of laccase was determined for 28-day duration at varying pH (3-10) and temperature (4-50 °C). The maximum storage stability was obtained at pH 4.5 and temperature 30 °C. Therefore, utilizing SMW for the degradation of OMPs from wastewater not only benefits in degradation but also reuses SMW agro waste, shedding light on agro waste management. Thus, SMW is a one-pot solution for both OMPs biodegradation and circularity in the economy.

Optimization of laccase enzyme extraction from spent mushroom waste of Pleurotus florida through ANN-PSO modeling: An ecofriendly and economical approach.

The cardinal focus of this study is to optimize the best reaction conditions for maximizing laccase activity from spent mushroom waste (SMW) of Pleurotus florida. Optimization process parameters were studied by the modeling techniques, artificial neural networking (ANN) embedded in particle swarm optimization (PSO), and response surface model (RSM). The best topology of ANN-PSO architecture was obtained on 4-10-1. The R2, IOA, MSE, and MAE values of the ANN model were obtained as 0.98785, 0.9939, 0.0023, and 0.0251 while, that of the RSM model were obtained as 0.74290, 0.9210, 0.0244, and 0.1110 respectively. The higher values of R2, IOA, and lower values of MSE and MAE of the ANN-PSO model depict that ANN-PSO outperformed compared to RSM and also verified the effectiveness of the ANN-PSO model. The ANN-PSO model performance demonstrates the robustness of the technique in optimizing laccase activity in SMW of P. florida. The optimization results revealed that pH 4.5, time 3 h, solid: solution ratio 1:5, and ABTS concentration of 1 mM was optimal for achieving maximum laccase activity at temperature 30 °C. The enzymatic activity of crude laccase enzyme was obtained as 1.185 U ml-1 without loss of enzyme activity. Additionally, crude laccase enzyme was 1.74 fold partially purified, and 83.54% of the enzyme was yielded. Out of all the independent process variables, ABTS and pH had an influence on laccase activity. Therefore, we anticipate that the findings of this investigation will reduce the ambiguity in maximizing laccase activity and ease the screening process. This study also highlights the comparative cost evaluation of crude laccase enzyme extracted from P. florida and commercial enzymes. There is a great potential for the utilization of the laccase enzyme extracted from SMW and using it for the degradation of recalcitrant micropollutants. Thus, SMW promises a cost-effective and sustainable approach leading towards circular economy.

Spent waste from edible mushrooms offers innovative strategies for the remediation of persistent organic micropollutants: A review.

Urgent and innovative strategies for removal of persistent organic micropollutants (OMPs) in soil, groundwater, and surface water are the need of the hour. OMPs detected in contaminated soils and effluents from wastewater treatment plants (WWTPs) are categorized as environmentally persistent pharmaceutical pollutants (EPPPs), and endocrine disrupting chemicals (EDCs), their admixture could cause serious ecological issues to the non-target species. As complete eradication of OMPs is not possible with the extant conventional WWTPs technology, the inordinate and reckless application of OMPs negatively impacts environmental regenerative and resilience capacity. Therefore, the cardinal focus of this review is the bioremediation of persistent OMPs through efficient application of an agro-waste, i.e. spent mushroom waste (SMW). This innovative, green, long-term strategy embedded in the circular economy, based on state of the art information is comprehensively assessed in this paper. SMW accrues ligninolytic enzymes such as laccase and peroxidase, with efficient mechanism to facilitate biodegradation of recalcitrant organic pollutants. It is vital in this context that future research should address immobilization of such enzymes to overcome quantitative and qualitative issues obstructing their widespread use in biodegradation. Therefore, dual benefit is gained from cultivating critical cash crops like mushrooms to meet the escalating demand for food resources and to aid in biodegradation. Hence, mushroom cultivation has positive environmental, social, and economic implications in developing countries like India.

Bacterial strains found in the soils of a municipal solid waste dumping site facilitated phosphate solubilization along with cadmium remediation.

Phosphate solubilizing bacteria (PSB) that can withstand high cadmium (Cd) stress is a desired combination for bioremediation. This study evaluated the Cd bioremediation potential of four PSB strains isolated from the contaminated soils of a municipal solid waste (MSW) discarding site (Guwahati, India). PSB strains were cultured in Pikovskaya (PVK) media, which led to higher acid phosphatase (ACP) activity and the release of organic acid. Optical density (OD) measurements were performed to determine the growth pattern of PSB; furthermore, Cd uptake by PSB was evaluated using infrared spectroscopy (IR) and X-Ray Diffraction (XRD) analyses. The 16S rRNA taxonomic analysis revealed that all the four promising PSB strains belonged to either Bacillus sp. or Enterobacter sp. One strain (SM_SS8) demonstrated higher tolerance towards Cd (up to 100 mg L-1). Flow cytometry analysis revealed 70.92%, 46.93% and 20.4% viability of SM_SS8 in 10, 50 and 100 mg L-1, respectively in PVK media containing Cd. This study has therefore substantiated the bioremediation of Cd from polluted soil by the PSB isolates. Thus, experimental results revealed a potential combo benefit, phosphate solubilization along with Cd remediation.

Premature Termination of MexR Leads to Overexpression of MexAB-OprM Efflux Pump in Pseudomonas aeruginosa in a Tertiary Referral Hospital in India.

OBJECTIVES: The present study was undertaken to investigate the mutations that are present in mexR gene of multidrug resistant (MDR) isolates of Pseudomonas aeruginosa collected from a tertiary referral hospital of north east India. METHODS: 76 MDR clinical isolates of P. aeruginosa were obtained from the patients who were admitted to or attended the clinics of Silchar medical college and hospital. They were screened phenotypically for the presence of efflux pump activity by an inhibitor based method. Acquired resistance mechanisms were detected by multiplex PCR. Real time PCR was performed to study the expression of mexA gene of MexAB-OprM efflux pump in isolates with increase efflux pump activity. mexR gene of the isolates with overexpressed MexAB-OprM efflux pump was amplified, sequenced and analysed. RESULTS: Out of 76 MDR isolates, 24 were found to exhibit efflux pump activity phenotypically against ciprofloxacin and meropenem. Acquired resistance mechanisms were absent in 11 of them and among those isolates, 8 of them overexpressed MexAB-OprM. All the 8 isolates possessed mutation in mexR gene. 11 transversions, 4 transitions, 2 deletion mutations and 2 insertion mutations were found in all the isolates. However, the most significant observation was the formation of a termination codon at 35th position which resulted in the termination of the polypeptide and leads to overexpression of the MexAB-OprM efflux pump. CONCLUSIONS: This study highlighted emergence of a novel mutation which is probably associated with multi drug resistance. Therefore, further investigations and actions are needed to prevent or at least hold back the expansion and emergence of newer mutations in nosocomial pathogens which may compromise future treatment options.