Rhizomucor miehei

A very attractive and efficient method for the synthesis of L-aminoacids via DKR has been reported by Turner et al. [41a,b]. They employed enzyme-catalyzed ring opening of 5(4H)-oxazolones in combination with a catalytic amount of Et3N. The relatively low pKa of the C-4 proton (8.9) of oxazolones facilitates racemization. Hydrolysis of the ester obtained through DKR, followed by debenzoylation, yields L-aminoacids in excellent enantiomeric excess (99.5%) (Figure 4.16). In their initial studies, they employed Rhizomucor miehei lipase (Lipozyme) as the biocatalyst [41]. More recently, they have obtained excellent results employing CALB [41bj. This method has also been employed by Bevinakatti [41c,d] and Sih [41e,fj. 

Lipase from Rhizomucor miehei Esterification of 1-butanol and oleic acid Oil-deionized water (96.6/3.4) 92.9 103... 

Lipase from Rhizomucor miehei Transesterification 2-ethyl-l-hexanol and rapeseed oil Oil-water (97/3) 93.8 103... 

The experiments were performed in a CINC V-02 separator also known as the CS-50 (15). Two Verder VL 500 control peristaltic tube pumps equipped with a double pump head (3,2 x 1,6 x 8R) were used to feed the CCS. In case of the enzymatic reaction, the low mix bottom plate was applied. To operate the reactor at a desired temperature, it was equipped with a jacket which was coimected to a temperature controlled water bath with an accuracy of 0.01°C. The CCS was fed with pure heptane and pure water, both with a flow rate of 6 mL/min. Subsequently, the centrifuge was started (40 Hz, which corresponds to 2400 rpm) and the set-up was allowed to equilibrate for a period of 1 h. At this point, the heptane feed stream was replaced by the organic feed stream (oleic acid (0.6M) and 1-bntanol (0.9M) in heptane). After equilibration for 10 minutes, the reaction in the CCS was started by replacing the water stream with the aqueous feed stream (0.1 M phosphate buffer pH 5.6 containing 1 g/1 of the lipase form Rhizomucor miehei). Samples were taken at regular intervals and analysed by GC. 

In a first experiment, the esterification of oleic acid with 1-butanol catalysed by a Rhizomucor miehei lipase was investigated (Scheme 4.3). Lipases usually function at the water/organic interface, which make them extremely suitable for use in the CCS. 

In this communication a study of the catalytic behavior of the immobilized Rhizomucor miehei lipase in the transesterification reaction to biodiesel production has been reported. The main drawbacks associated to the current biodiesel production by basic homogeneous catalysis could be overcome by using immobilized lipases. Immobilization by adsorption and entrapment have been used as methods to prepare the heterogeneous biocatalyst. Zeolites and related materials have been used as inorganic lipase supports. To promote the enzyme adsorption, the surface of the supports have been functionalized by synthesis procedures or by post-treatments. While, the enzyme entrapping procedure has been carried out by sol-gel method in order to obtain the biocatalyst protected by a mesoporous matrix and to reduce its leaching after several catalytic uses. 

A major cause of suboptimal activity in organic solvent results from the removal of essential water during enzyme dehydration. All enzymes require some water in order to retain activity through the provision of conformational flexibihty. Particularly in the case of lipases, the amount of water can be so low that it appears that none is required. For example, following the development of suitable techniques to analyse low water concentrations, it has been reported that the lipase from Rhizomucor miehei retains 30 % of its optimum activity with as little as two or three water molecules per molecule of enzyme.Owing to the apparent absence of water in some exceptional cases, the term biocatalysis in anhydrous solvent is commonly used, although in the vast majority of cases a monolayer of water is required for optimal activity (although this is often stUl well below its solubility limit in water-immiscible solvent). ... 

A lipase from Rhizomucor miehei is immobilized onto a porous polyamide support with a bead diameter of 0.3 mm and used for the hydrolysis of 100 mM caprylin at pH 7.0 and 30°C, in a continous plug flow reactor. Calculate the amount of immobilized lipase needed to achieve a conversion degree of 97% with an inlet flow rate of 1 l.min ... 

Other microbial lipases have also been successfully used in anhydrous ionic liquids, e.g., from Alcaligenes sp. (AsL) [54, 58], CaLA, Rhizomucor miehei lipase (RmL), and Thermomyces lanuginosus lipase (TIL) [54]. The lipase from pig pancreas (porcine pancreas lipase, PPL), the only mammalian lipase that has been subjected to ionic liquids, catalyzed transesterificationin[BMIm][NTf2]butnotin[BMIm][PF6]... 

Tajdini, F., Ali Amini, M., Nafissi-Varcheh, N., and Ali Faramarzi, M. (2010). Production, physiochemical and antimicrobial properties of fungal chitosan from Rhizomucor miehei and Mucor racemosus. Int. J. Biol. Macromol. 47,180-183. 

Rhizomucor miehei secretes a lipase that is reported to give satisfactory results in Italian cheese manufacture. This enzyme has been characterized and is commercially available as Piccantase . Lipases secreted by selected strains of Penicillium roqueforti and P. candidum are considered to be potentially useful for the manufacture of Italian and other cheese varieties. 

The annual production would be 550 t of oleyl oleate with market price 5 EURO/kg. As biocatalyst, an immobilized Rhizomucor miehei lipase - Lipozyme IM - product from NOVO Nordisk was used. 

Liew, M., Ghazali, H., Long, K., Lai, O., Yazid, A. 2001. Physical properties of palm kernel olein-anhydrous milk fat mixtures transesterified using mycelium-bound lipase from Rhizomucor miehei. Food Chem. 72, 447 454. 

Safari et al. (1993) examined the interesterification of milk fat by the lipase from Rhizomucor miehei in various organic solvents (hexane, hexane-choloroform (70 30, v/v), and hexane-ethylacetate (70 30, v/v)). The addition of chloroform or ethyl acetate to hexane increased lipase activity. It was suggested that the polarity of the solvent influences the partitioning of water in the system with consequent effects on enzymic activity. Bornaz et al. 

Interesterification of blends of milk fat and palm kernel olein by a mycelium-bound lipase from Rhizomucor miehei or a commercially immobilized enzyme preparation resulted in a lower slip melting point and solid fat content. An interesterified product made from a 70 30 mixture of palm kernel olein and anhydrous milk fat was considered to be suitable for use in ice cream (Liew et al., 2001). 

Maruyama, K., Shimada, Y., Baba,T., Ooguri,T., Sugihara, A.,Tominaga, Y., and Mori-yama, S. 2000. Purification of ethyl docosahexaenoate through selective alcoholysis with immobilized Rhizomucor miehei lipase. J. Jpn. Oil Chem. Soc., 49, 793-799. 

In addition to the recombinant chymosin, several coagulants for cheese-making are available from microbial origin. These are endopro-teases from Rhizomucor miehei, Rhizomucor pussilus, and incidentally the plant-derived endothiapepsin from Cryphonectria parasitica. 

Lipase [Rhizomucor (Mucor) miehei] Produced as an off white to tan powder or as a liquid by controlled fermentation using Rhizomucor miehei. Soluble in water (the solution is usually light yellow to dark brown), but practically insoluble in alcohol, in chloroform, and in ether. Major active principle lipase. Typical applications used in the hydrolysis of lipids, in the manufacture of cheese, and in the removal of haze in fruit juices. 

Aspergillus oryzae var. (4) Aspergillus niger var. (5) Rhizomucor miehei (6) Candida rugosa ... 

Rennet protease (1) fourth stomach of ruminant animals (2) Endothia parasitica (3) Rhizomucor miehei (4) Rhizomucor pusillus (Lindt) (5) Bacillus cereus none 3.4.23.1 3.4.23.4 3.4.23.22 3.4.23.23... 

The enzymatic esterification of oleic acid and oleyl alcohol to obtain oleyl oleate, which is a synthetic analogue of jojoba oil, was studied. The reaction was catalyzed by a commercially available immobilized lipase from Rhizomucor miehei. As solvents, carbon dioxide and liquid n-butane were used. Reactions were performed in a batch and in continuously operating high pressure reactors. 

Enzyme preparation. The enzyme preparation Lipozyme IM, which is a Rhizomucor miehei lipase, immobilized on a macroporous anion exchange resin, was kindly donated from NOVO Nordisk AS (Copenhagen, Denmark). 

Reaction performance. Supercritical carbon dioxide was used as a reaction media for the enzymatic synthesis of oleyl oleate directly from oleic acid and oleyl alcohol. Reaction was catalyzed by immobilized lipase from Rhizomucor miehei-Ltpozyme IM. Reactions were carried out in the high pressure batch and continuous reactor. 

Esters of fatty acids with monohydric alcohols find applications as emollients in cosmetics. They are prepared by acid- or base-catalyzed (trans)esterifications [200, 205]. As with biodiesel production, the use of enzymatic catalysis offers potential benefits but in the case of these specialty fatty acid esters there is a special advantage the products can be labelled as natural. Consequently, they command a higher price in personal care products where natural is an important customer-perceived advantage. Examples include the synthesis of isopropylmy-ristate by CaLB-catalyzed esterification [206] and n-hexyl laurate by Rhizomucor miehei lipase (Lipozyme IM-77)-catalyzed esterification [207] (see Fig. 8.38). 

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