Investigation of Halohydrins Degradation by Whole Cells and Cell-free Extract of Pseudomonas putida DSM 437: A Kinetic Approach

The biodegradation of two halohydrins (1,3-dichloro-2-propanol and 3-chloro-1,2-propanediol) by P. putida DSM 437 was investigated. Intact cells of previously acclimatized P. putida DSM 437 as well as cell-free extracts were used in order to study the degradation kinetics. When whole cells were used, a maximum biodegradation rate of 3-CPD (vmax= 1.28.10–5 mmol mg–1 DCW h–1) was determined, which was more than 4 times higher than that of 1,3-DCP. However, the affinity towards both halohydrins (Km) was practically the same. When using cell-free extract, the apparent vmax and Km values for 1,3-DCP were estimated at 9.61.10–6 mmol mg–1 protein h–1 and 8.00 mM, respectively, while for 3-CPD the corresponding values were 2.42.10–5 mmol mg–1 protein h–1 and 9.07 mM. GC-MS analysis of cell-free extracts samples spiked with 1,3-DCP revealed the presence of 3-CPD and glycerol, intermediates of 1,3-DCP degradation pathway. 3-CPD degradation was strongly inhibited by the presence of epichlorohydrin and to a lesser extent by glycidol, intermediates of dehalogenation pathway. This work is licensed under a Creative Commons Attribution 4.0 International License

Application of Different Processes for the Biodegradation of 1,3-dichloro-2-propanol by the Bacterium Pseudomonas putidaDSM437

1,3-Dichloro-2-propanol (1,3-DCP), is a highly toxic compound used in many industrial processes. Biodegradation of 1,3-DCP, by the bacterial strain Pseudomonas putida DSM 347, was studied applying three different processes. A number of combinations, with respect to glucose and 1,3-DCP concentration were examined during batch process. When the initial concentration of 1,3-DCP was 600 mg L–1 in the presence of 400 mg L–1 glucose, the biodegradation degree and rate were 10.8 % and 0.68 mg L–1 h–1 respectively. 1,3-DCP biodegradation by the resting cells of P. putida DSM 347 was tested at mass concentrations from = 200 to 1 000 mg L–1 using biomass concentration of 5 g dry cell mass L–1. Biodegradation of 1,3-DCP ranged from 84 to 90 %, initial biodegradation rates ranged from r = 2.36 to 10.55 mg L–1 h–1, while dependence of both parameters from the initial concentration of halohydrin was observed. A system of two Continuous Stirred Tank Reactors (CSTRs) in series was developed for the biodegradation of a highly toxic stream of 1,3-DCP (2000 mg L–1). The overall biodegradation degree of the system was 68 %, while biodegradation rates of the first and second bioreactor were r = 2.88 and 5.21 mg L–1 h–1 respectively

Mastitis and Mammary Abscess Management Audit (MAMMA) in the UK and Ireland

BACKGROUND: The aim of this multicentre prospective audit was to describe the current practice in the management of mastitis and breast abscesses in the UK and Ireland, with a specific focus on rates of surgical intervention. METHODS: This audit was conducted in two phases from August 2020 to August 2021; a phase 1 practice survey and a phase 2 prospective audit. Primary outcome measurements for phase 2 included patient management pathway characteristics and treatment type (medical/radiological/surgical). RESULTS: A total of 69 hospitals participated in phase 2 (1312 patients). The key findings were a high overall rate of incision and drainage (21.0 per cent) and a lower than anticipated proportion of ultrasound-guided aspiration of breast abscesses (61.0 per cent). Significant variations were observed regarding the rate of incision and drainage (range 0-100 per cent; P \u3c 0.001) and the rate of needle aspiration (range 12.5-100 per cent; P \u3c 0.001) between individual units. Overall, 22.5 per cent of patients were admitted for inpatient treatment, out of whom which 72.9 per cent were commenced on intravenous antibiotics. The odds of undergoing incision and drainage for a breast abscess or being admitted for inpatient treatment were significantly higher if patients presented at the weekend compared with a weekday (P ≤ 0.023). Breast specialists reviewed 40.9 per cent of all patients directly, despite the majority of patients (74.2 per cent) presenting within working hours on weekdays. CONCLUSIONS: Variation in practice exists in the management of mastitis and breast abscesses, with high rates of incision and drainage in certain regions of the UK. There is an urgent need for a national best-practice toolbox to minimize practice variation and standardize patient care

Medium composition overturns the widely accepted sulfate-dependent repression of desulfurization phenotype in Rhodococcus qingshengii IGTS8

Microbial desulfurization has been extensively studied as a promising alternative to the widely applied chemical desulfurization process. Sulfur removal from petroleum and its products becomes essential, as the environmental regulations become increasingly stringent. Rhodococcus qingshengii IGTS8 has gained ground as a naturally occurring model biocatalyst, due to its superior specific activity for desulfurization of dibenzothiophene (DBT). Recalcitrant organic sulfur compounds—DBT included—are preferentially removed by selective carbon-sulfur bond cleavage to avoid a reduction in the calorific value of the fuel. The process, however, still has not reached economically sustainable levels, as certain limitations have been identified. One of those bottlenecks is the repression of catalytic activity caused by ubiquitous sulfur sources such as inorganic sulfate, methionine, or cysteine. Herein, we report an optimized culture medium for wild-type stain IGTS8 that completely alleviates the sulfate-mediated repression of biodesulfurization activity without modification of the natural biocatalyst. Medium C not only promotes growth in the presence of several sulfur sources, including DBT, but also enhances biodesulfurization of resting cells grown in the presence of up to 5 mM sulfate. Based on the above, the present work can be considered as a step towards the development of a more viable commercial biodesulfurization process. © 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC

Biochemical and catalytic properties of two intracellular β-glucosidases from the fungus Penicillium decumbens active on flavonoid glucosides

In the presence of rutin as sole carbon source, Penicillium decumbens produces two intracellular β-glucosidases named GI and G II, with molecular masses of 56,000 and 460,000Da, respectively. The two proteins have been purified to homogeneity. GI and G II composed of two and four equal sub-units, respectively and displayed optimal activity at pH 7.0 and temperature 65-75°C. Both β-glucosidases were competitively inhibited by glucose and glucono-δ-lactone. GI and GII exhibited broad substrate specificity, since they hydrolyzed a range of (1,3)-, (1,4)- and (1,6)-β-glucosides as well as aryl β-glucosides. Determination of kcat/Km revealed that GII hydrolyzed 3-8 times more efficiently the above-mentioned substrates. The ability of GI and GII to deglycosylate various flavonoid glycosides was also investigated. Both enzymes were active against flavonoids glycosylated at the 7 position but GII hydrolyzed them 5 times more efficiently than G I. Of the flavanols tested, both enzymes were incapable of hydrolyzing quercetrin and kaempferol-3-glucoside. The main difference between GI and GII as far as the hydrolysis of flavanols is concerned, was the ability of GII to hydrolyze the quercetin-3-glucoside. © 2003 Elsevier B.V. All rights reserved

Fungal multienzyme production on industrial by-products of the citrus-processing industry

Orange peels is the principal solid by-product of the citrus processing industry and the disposal of the fresh peels is becoming a major problem to many factories. Dry citrus peels are rich in pectin, cellulose and hemicellulose and may be used as a fermentation substrate. Production of multienzyme preparations containing pectinolytic, cellulolytic and xylanolytic enzymes by the mesophilic fungi Aspergillus niger BTL, Fusarium oxysporum F3, Neurospora crassa DSM 1129 and Penicillium decumbens under solid-state fermentation (SSF) on dry orange peels was enhanced by optimization of initial pH of the culture medium and initial moisture level. Under optimal conditions A. niger BTL was by far the most potent strain in polygalacturonase and pectate lyase, production followed by F. oxysporum F3, N. crassa DSM 1129 and P. decumbens. N. crassa DSM 1129 produced the highest endoglucanase activity and P. decumbens the lowest one. Comparison of xylanase production revealed that A. niger BTL produced the highest activity followed by N. crassa DSM 1129, P. decumbens and F. oxysporum F3. N. crassa DSM 1129 and P. decumbens did not produce any β-xylosidase activity, while A. niger BTL produced approximately 10 times more β-xylosidase than F. oxysporum F3. The highest invertase activity was produced by A. niger BTL while the lowest ones by F. oxysporum F3 and P. decumbens. After SSF of the four fungi, under optimal conditions, the fermented substrate was either directly exposed to autohydrolysis or new material was added, and the in situ produced multienzyme systems were successfully used for the partial degradation of orange peels polysaccharides and the liberation of fermentable sugars.</p