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Candida rugosa

Manufactured by Merck Group
Sourced in Germany

Candida rugosa is a type of laboratory equipment used for various biochemical and biotechnological applications. It serves as a source of lipase, an enzyme that catalyzes the hydrolysis of fats and oils. The enzyme's core function is to facilitate the breakdown of lipids, which is a crucial process in many industrial and research settings.

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5 protocols using candida rugosa

1

Enzymatic Hydrolysis of Sea Buckthorn Pulp Oil

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Sea buckthorn pulp oil (obtained from Henry Lamotte Oils GmbH, Bremen, Germany) was enzymatically hydrolyzed. Therefore, 5 mg oil was dispersed in 2 mL phosphate buffer (50 mM, pH 7.5) by means of an ultrasonic bath for 5 min. Lipase (6 U, E.C. 3.1.1.3) from Candida rugosa (Sigma Aldrich) was added and the mixture was incubated in a rotary shaker (40 rpm) at 24 °C for 3 h in the dark.
Besides the hydrolyzed lipid extract, standard fatty acids palmitic, palmitoleic, oleic, and linoleic acid were prepared as indicated above. Biotransformation experiments were executed under optimized conditions. 150 U/L Csα-DOX and 18.75 U/L VhFALDH were added to the substrate-cofactor dispersion. After incubation for 4 h, the reaction was stopped by addition of HCl as described above, and the samples were stored at –20 °C until analysis.
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2

Lipase-catalyzed Fatty Acid Oxidation

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High-quality (>99%) ALA and linoleic acid (LA; 18:2n-6) were purchased from Nu-Check Prep (Elysian, MN) and Sigma (St. Louis, MO, USA), respectively. 9-Hydroperoxy-trioctadecatrienoic acid (9-HPOTE) and 9S-HOTE standards were purchased from Cayman Chemicals (Ann Arbor, MI). 9R-HOTE standard was prepared by the previously reported procedure [23 ]. Solvents, including 2-propanol, acetonitrile, n-hexane, and methanol, were high-performance liquid chromatography (HPLC) grade and ethanol and ethyl acetate for extraction were reagent grade. These solvents were supplied from Duksan Chemical (Ansan, Korea). Commercial PO was obtained from a local market. Luria-Bertani (LB) medium was purchased from Fermentas (Vilnius, Lithuania). Ampicillin, boric acid, and isopropyl-β-d-thiogalactopyranoside (IPTG) were purchased from USB (Cleveland, OH, USA). Lipases from recombinant Aspergillus oryzae (AOL), Candida rugosa (CRL), Pseudomonas cepacia (PCL), Pseudomonas fluorescense (PFL), Rhizomucor meihei (RML), and Thermomyces lanoginosus (TLL) were obtained from Sigma.
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3

Enzymatic Degradation of Reeds with PBAT

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PBAT (TH801T) was purchased from Xinjiang Blue Ridge Tunhe Polyester Co., Ltd (Changji, China). It has a specific gravity of 1.21 g/cm3 and a melt mass flow rate (MFR) of 5.0 g/10 min (190 °C), with a number-average molecular weight of 6.12 × 104 g/mol. Reeds were collected from the shore of the lake (Baiyangdian, Baoding, China). For enzymatic degradation, the following enzymes and reagents were used: Proteinase K (≥30 units/mg) from Tritirachium album (Blirt, Gdansk, Poland), esterase (≥10 units/mg) from Bacillus subtilis (Sigma-Aldrich, Bavaria, Germany), lipase (15–25 units/mg) from Candida rugosa (Sigma-Aldrich, Bavaria, Germany), and cellulase (≥200 FPU/mL) from Trichoderma reesei (Novozymes A/S, Copenhagen, Denmark). Tris(hydroxymethyl)aminomethane-HCl (0.1 M, pH = 8.0, Sigma-Aldrich, Germany) and citric acid-sodium citrate (0.1 M, pH = 4.8, Sigma-Aldrich, Germany) were used as buffers for enzyme degradation, sodium azide (NaN3) (Sigma Chemical Co., St. Louis, MI, USA) was used as an antifungal agent, and calcium chloride (Sigma Chemical Co., USA) was used as an activator of enzyme degradation. Other chemical reagents were obtained from Sinopharm Chemical Reagent (Shanghai, China).
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4

Hydrolysis of Diverse Vegetable Oils

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Hydrolysis reactions were performed by Aspergillus niger (product number 62301), Candida rugosa (product number 62316), Rhizopus oryzae (product number 62305), Rhizopus niveus (product number 62310) and Rhizomucor miehei (formerly known as Mucor miehei, product number 62298) free lipases obtained from Sigma–Aldrich (Munich, Germany). Seven oils, i.e., olive (refined; product number O1514), linseed (product number 430021), peanut (product number P2144), grape seed (product number W233218), rapeseed (product number 83450), almond (product number 63445) and menhaden fish (refined; product number F8020) oils, were involved to the assay. All oils were purchased from Sigma–Aldrich (Munich, Germany). PA (5.30–25.23%) was a major saturated fatty acid component in the oils used as determined by gas chromatography (GC) [46 (link)]. Concerning unsaturated fatty acids, OA (5.12–80.05%) and LA (1.91–23.79%) were among the major components of the oils. In addition, ALA (58.33%) in linseed oil and EPA (12.11%) and DHA (5.47%) in menhaden fish oil were also predominant fatty acid components. Individual fatty acids, i.e., EPA and ALA, were obtained from VWR (Debrecen, Hungary).
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5

Fungal Enzyme Production in Liquid Medium

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The fungi were grown in Erlenmeyer flasks containing 100 ml of liquid medium, as described by Nascimento et al. (2014) . The medium was composed of NH2NO3 (1.0 g/l), MgSO4.7H2O (0.6 g/l), KH2PO4 (1.0 g/l), peptone (20 g/l) and olive oil (1% and 0.065%), pH 6. 100 µl of spore suspension was inoculated (10 6 spores/ml) under sterile conditions and incubated in a shaker at 28°C, 160 rpm for 7 days. Every 24 h, 1 ml aliquots were taken and filtered for subsequent measurement of enzyme activity. Cultivations were performed in triplicate. The commercial enzyme produced by Candida rugosa (Sigma Aldrich) was used as a standard for comparative purposes.
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