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Adsorption of lipase on hollow fiber membrane chips

Shamel MM, Azaha RB, Al-Zuhair S

Artif Cells Blood Substit Immobil Biotechnol, 2005;33(4):423-33.
PMID: 16317961

The amount of lipase from Mucor miehei adsorption on ultrafiltration polysulfone hollow fiber membrane chips has been determined using different lipase concentrations at three different temperatures, namely 30, 35, and 40 degrees C. It was experimentally shown that adsorption of lipase increases with temperature. The results were used to evaluate the constants found in the Langmuir adsorption isotherm model coupled with the Van't Hoff's relationship. A temperature dependence correlation for the amount of adsorbed lipase activity, alip,ads, and that present in the supernatant solution, alip,free was determined. The effect of varying the concentration on a cross-linking agent, namely, glutaraldehyde, to the membrane chips was also tested. It was found that, under the same operating conditions, the amount of lipase adsorbed on polysulfone membranes was increased dramatically after pre-treating the membrane with 1% Glutaraldehyde. However, increasing the concentration of the cross-linking agent has a low effect on the amount of lipase adsorbed.

Matched MeSH terms: Mucor/enzymology
Fulltext Role of Snf-β in lipid accumulation in the high lipid-producing fungus Mucor circinelloides WJ11

Nosheen S, Naz T, Yang J, Hussain SA, Fazili ABA, Nazir Y, et al.

Microb Cell Fact, 2021 Feb 27;20(1):52.
PMID: 33639948 DOI: 10.1186/s12934-021-01545-y

BACKGROUND: Mucor circinelloides WJ11 is a high-lipid producing strain and an excellent producer of γ-linolenic acid (GLA) which is crucial for human health. We have previously identified genes that encode for AMP-activated protein kinase (AMPK) complex in M. circinelloides which is an important regulator for lipid accumulation. Comparative transcriptional analysis between the high and low lipid-producing strains of M. circinelloides showed a direct correlation in the transcriptional level of AMPK genes with lipid metabolism. Thus, the role of Snf-β, which encodes for β subunit of AMPK complex, in lipid accumulation of the WJ11 strain was evaluated in the present study.
RESULTS: The results showed that lipid content of cell dry weight in Snf-β knockout strain was increased by 32 % (from 19 to 25 %). However, in Snf-β overexpressing strain, lipid content of cell dry weight was decreased about 25 % (from 19 to 14.2 %) compared to the control strain. Total fatty acid analysis revealed that the expression of the Snf-β gene did not significantly affect the fatty acid composition of the strains. However, GLA content in biomass was increased from 2.5 % in control strain to 3.3 % in Snf-β knockout strain due to increased lipid accumulation and decreased to 1.83 % in Snf-β overexpressing strain. AMPK is known to inactivate acetyl-CoA carboxylase (ACC) which catalyzes the rate-limiting step in lipid synthesis. Snf-β manipulation also altered the expression level of the ACC1 gene which may indicate that Snf-β control lipid metabolism by regulating ACC1 gene.
CONCLUSIONS: Our results suggested that Snf-β gene plays an important role in regulating lipid accumulation in M. circinelloides WJ11. Moreover, it will be interesting to evaluate the potential of other key subunits of AMPK related to lipid metabolism. Better insight can show us the way to manipulate these subunits effectively for upscaling the lipid production. Up to our knowledge, it is the first study to investigate the role of Snf-β in lipid accumulation in M. circinelloides.

Matched MeSH terms: Mucor/metabolism*
Vertical distribution of mycostasis in Malayan soils

Griffiths DA

Can J Microbiol, 1966 Feb;12(1):149-63.
PMID: 5923132
Matched MeSH terms: Mucor*
Biochemical events leading to the diversion of carbon into storage lipids in the oleaginous fungi Mucor circinelloides and Mortierella alpina

Wynn JP, Hamid AA, Li Y, Ratledge C

Microbiology (Reading), 2001 Oct;147(Pt 10):2857-2864.
PMID: 11577164 DOI: 10.1099/00221287-147-10-2857

The biochemical events associated with the onset of lipid accumulation in Mucor circinelloides and Mortierella alpina, under conditions of nitrogen-limited growth, have been elucidated; they differ in key aspects from those described in oleaginous yeasts. The NAD+:isocitrate dehydrogenases of Mc. circinelloides and Mort. alpina were not absolutely dependent on AMP for activity. Furthermore, changes in the cellular adenine nucleotide pools and energy charge were different from those reported for oleaginous yeasts. In Mc. circinelloides ATP, ADP and AMP concentrations all decreased by 50% after nitrogen limitation, leading to a constant energy charge at the expense of the size of the total adenylate pool. Pyruvate carboxylase in Mc. circinelloides was cytosolic, having implications for the organization of lipid synthesis in filamentous fungi. As a result of the data obtained, a revised and more concerted mechanism for the initiation of storage lipid accumulation is put forward for filamentous fungi.

Matched MeSH terms: Mucor/growth & development; Mucor/metabolism*
Fulltext Coculturing of Mucor plumbeus and Bacillus subtilis bacterium as an efficient fermentation strategy to enhance fungal lipid and gamma-linolenic acid (GLA) production

Mohamed H, Awad MF, Shah AM, Sadaqat B, Nazir Y, Naz T, et al.

Sci Rep, 2022 Jul 30;12(1):13111.
PMID: 35908106 DOI: 10.1038/s41598-022-17442-2

This study aimed to improve lipid and gamma-linolenic acid (GLA) production of an oleaginous fungus, Mucor plumbeus, through coculturing with Bacillus subtilis bacteria, optimising the environmental and nutritional culture conditions, and scaling them for batch fermentation. The maximum levels of biomass, lipid, fatty acid, and GLA in a 5 L bioreactor containing cellobiose and ammonium sulfate as the optimal carbon and nitrogen sources, respectively, achieved during the coculturing processes were 14.5 ± 0.4 g/L, 41.5 ± 1.3, 24 ± 0.8, and 20 ± 0.5%, respectively. This strategy uses cellobiose in place of glucose, decreasing production costs. The nutritional and abiotic factor results suggest that the highest production efficiency is achieved at 6.5 pH, 30 °C temperature, 10% (v/v) inoculum composition, 200 rpm agitation speed, and a 5-day incubation period. Interestingly, the GLA concentration of cocultures (20.0 ± 0.5%) was twofold higher than that of monocultures (8.27 ± 0.11%). More importantly, the GC chromatograms of cocultures indicated the presence of one additional peak corresponding to decanoic acid (5.32 ± 0.20%) that is absent in monocultures, indicating activation of silent gene clusters via cocultivation with bacteria. This study is the first to show that coculturing of Mucor plumbeus with Bacillus subtilis is a promising strategy with industrialisation potential for the production of GLA-rich microbial lipids and prospective biosynthesis of new products.

Matched MeSH terms: Mucor
Mycoremediation--a prospective environmental friendly technique of bioseparation and dewatering of domestic wastewater sludge

Molla AH, Fakhru'l-Razi A

Environ Sci Pollut Res Int, 2012 Jun;19(5):1612-9.
PMID: 22134862 DOI: 10.1007/s11356-011-0676-0

INTRODUCTION: Environmental safe and friendly management and disposal of wastewater sludge is a problem of every treatment plant throughout the world. Bioseparation and dewaterability of raw domestic wastewater sludge were evaluated for proper management and disposal by mycoremediation, i.e., using prior grown 2% (v/v) spore suspension of filamentous fungal (Mucor hiemalis Wehmer) broth inoculation, which were grown in 2% (w/v) solution of malt extract and wheat flour for 48-60 h in orbital shaker.
DISCUSSION: Within 2-3 days of treatment application, encouraging results were achieved in total dry solids (TDS), total suspended solid (TSS), turbidity, chemical oxygen demand (COD), specific resistance to filtration (SRF), and pH due to fungal treatment in recognition of bioseparation and dewaterability of wastewater sludge compared to control. The significant reduction of TDS was remarked at fungal biomass (FB) in wheat flour (WF) treatment. The removal of TSS, turbidity, COD, and SRF were observed 96.0%, 99.4%, 92.6%, and 97.6%, respectively, in supernatant at 5 days by FB in WF. The SRF measuring the dewaterability was decreased with maximum (0.26 × 10(-12) mg/kg) equivalent to 95.5% at 2 days in FB in WF also. FB in WF broth is a potential, environmental friendly, comparatively low-cost biological technique which might play the significant role for bioremediation and bioseparation of domestic wastewater sludge. The present technique may bring a dynamic change in treatment of wastewater in future.

Matched MeSH terms: Mucor*
Fulltext Redirecting Metabolic Flux towards the Mevalonate Pathway for Enhanced β-Carotene Production in M. circinelloides CBS 277.49

Naz T, Nazir Y, Nosheen S, Ullah S, Halim H, Fazili ABA, et al.

Biomed Res Int, 2020;2020:8890269.
PMID: 33457420 DOI: 10.1155/2020/8890269

Carotenoids produced by microbial sources are of industrial and medicinal importance due to their antioxidant and anticancer properties. In the current study, optimization of β-carotene production in M. circinelloides strain 277.49 was achieved using response surface methodology (RSM). Cerulenin and ketoconazole were used to inhibit fatty acids and the sterol biosynthesis pathway, respectively, in order to enhance β-carotene production by diverting metabolic pool towards the mevalonate pathway. All three variables used in screening experiments were found to be significant for the production of β-carotene. The synergistic effect of the C/N ratio, cerulenin, and ketoconazole was further evaluated and optimized for superior β-carotene production using central composite design of RSM. Our results found that the synergistic combination of C/N ratios, cerulenin, and ketoconazole at different concentrations affected the β-carotene productions significantly. The optimal production medium (std. order 11) composed of C/N 25, 10 μg/mL cerulenin, and 150 mg/L ketoconazole, producing maximum β-carotene of 4.26 mg/L (0.43 mg/g) which was 157% greater in comparison to unoptimized medium (1.68 mg/L, 0.17 mg/g). So, it was concluded that metabolic flux had been successfully redirected towards the mevalonate pathway for enhanced β-carotene production in CBS 277.49.

Matched MeSH terms: Mucor*
Fulltext Extraction, purification and characterisation of a milk-clotting protease from ‘kesinai’ (Streblus asper Lour.) leaves

Pagthinathan, M., Ghazali, H.M., Yazid, A.M., Foo, H.L.

International Food Research Journal, 2019;26(3):913-922.
MyJurnal

Extracts from ‘kesinai’ (Streblus asper) leaves were investigated as a potential source of enzymes that can serve as an alternative to calf rennet in cheese making. Different types of extraction buffers were investigated namely sodium acetate buffer (pH 4.2-5.0), phosphate buffer (pH 6.0-7.0) and Tris-HCl buffer (pH 7.0-9.0). Finally, the milk-clotting enzyme was extracted using 100 mM Tris-HCl buffer (pH 7.4) with and without 5.0 mg/mL polyvinylpyrrolidone, 0.015 mL/mL Triton X-100 and 2 mM sodium metabisulphite. Purification was carried out using acetone precipitation, and ion-exchange and size-exclusion chromatographic techniques. Results showed that 100 mM Tris-HCl buffer (pH 7.4) was the most efficient extraction buffer among the buffers used in the extraction study. After the final purification step of size-exclusion chromatography, the enzyme was purified 3.3-fold with 42.3% of recovery. The enzyme showed an optimum temperature and pH at 60°C and pH 7.4, respectively. The enzyme was stable up to 70°C for one hour and the partially purified enzyme retained 83% and 96% of its original activity at pH 6.0 and 8.0, respectively. The molecular weight of the partially enzyme was estimated to be 75.8 kDa on SDS-PAGE. The milk-clotting activity of ‘kesinai’ enzyme was found to be lower than that of commercial Mucor rennet.

Matched MeSH terms: Mucor