Novozymes

Hexanediol (0.5 mol/L), dimethyl fumarate (0.5 mol/L), toluene and Novozyme (33.3 g/L) are introduced in a thermostatted double-jacketted reactor fitted with a short thermostatted distillation column and a nitrogen inlet. The temperature is set at 60°C and a nitrogen flow (0.2 L/min) is bubbled into reaction medium. Methanol and toluene are collected in a flask and die volume of the solution is held constant by addition of toluene. After reaction (15 days) the catalyst is removed by filtration and the solvent is evaporated under reduced... 

For desymmetrization of diesters 3 via their hydrolysis in water, pig Hver esterase [12], o -chymotrypsin [12, 13a], and Candida antarctica Hpase (CAL-B) [14] were successfully used. However, further studies showed that respective anhydrides 5 can be used as substrates for enzyme-catalyzed desymmetrization in organic solvents [15]. The desired monoesters 4 were obtained in high yield in this way, using immobilized enzymes Novozym 435 or Chirazyme L-2 (Scheme 5.3). After the reaction, enzymes were filtered off, organic solvents were evaporated, and the crude products were crystalHzed. This was a much simpler experimental procedure in which control of the reaction progress was not necessary, and aU problems associated with extraction of products from aqueous phase and their further purification were omitted [15]. 

Defatted soy flour was suspended in water to 8% (v/w) protein, pH was adjusted to 6 by HCl before 640 mg/L Novozyme 415 (a-galactosidase), 10 mg/L Phytase L (Novo Nordisk) and 53 mg enzyme protein/L rhamnogalacturonase B were added. After 4 hours at 50 C the pH was adjusted to 8 by NaOH, and the slurry was centrifuged. The supernatant was pasteurized (85°C, 5 minutes) and freezedried. Protein was measured as 6.25 x Kjeldahl N. Phytate was measured as described in [16], and dietary fibres were analysed as described in [17]. 

The catalysts was added after the reactants were fed in the tank reactor and pressure and temperature were set to the target values [84]. The study was performed using an immobilized lipase, Novozym-435 , as biocatalyst. The temperature was set to 65-75 °C and the pressure was reduced (60 mmHg). A catalyst concentration of 1-5% with an acid alcohol ratio of 1 3, 1 1 or 3 1 was used. 

The ability of enzymes to achieve the selective esterification of one enantiomer of an alcohol over the other has been exploited by coupling this process with the in situ metal-catalysed racemisation of the unreactive enantiomer. Marr and co-workers have used the rhodium and iridium NHC complexes 44 and 45 to racemise the unreacted enantiomer of substrate 7 [17]. In combination with a lipase enzyme (Novozyme 435), excellent enantioselectivities were obtained in the acetylation of alcohol 7 to give the ester product 43 (Scheme 11.11). A related dynamic kinetic resolution has been reported by Corberdn and Peris [18]. hi their chemistry, the aldehyde 46 is readily racemised and the iridium NHC catalyst 35 catalyses the reversible reduction of aldehyde 46 to give an alcohol which is acylated by an enzyme to give the ester 47 in reasonable enantiomeric excess. 

Phytic acid (inisitol hexakisphosphate) is the main storage form of phosphorus in plants. The phosphorus is not bioavailable to non-ruminants as they lack the enzymes to break it down. Novozyme has developed a commercial enzyme, phytase, that can be added to animal feed to release the phosphorus. No inorganic phosphorus needs to be added. This shift in the source of phosphorous has a large impact on the environmental footprint of pig farming. 

Chemo-enzymatic epoxidation of unsaturated fatty acids with aqueous H2O2 has been conducted with considerable success and here we have a remarkable situation that undesirable ring opening of the epoxide does not occur. Excellent activity and stability has been realized with Novozym 435, a Candida antartica lipase B immobilized on polyacryl. This enzyme is readily separable, can be used several times without loss of activity, and has a turnover of more than 2,00,000 moles of products per mole of catalyst (Bierman et al., 2000). 

Novozymes is a market leader in enzyme solutions. Their manufacturing capabilities are based on an advanced biotech platform, for identifying new enzyme applications. Novozymes produces and sells more than 500 enzyme products covering more than 20 different industries in the food, feed and technical sectors, in 120 countries. With more than 75 types of enzymes and almost 600 different products, Novozymes has the world largest enzymes portfolio. 

In conventional synthetic transformations, enzymes are normally used in aqueous or organic solvent at moderate temperatures to preserve the activity of enzymes. Consequently, some of these reactions require longer reaction times. In view of the newer developments wherein enzymes can be immobilized on solid supports [183], they are amenable to relatively higher temperature reaction with adequate pH control. The application of MW irradiation has been explored with two enzyme systems namely Pseudomonas lipase dispersed in Hyflo Super Cell and commercially available SP 435 Novozym (Candida antarctica lipase grafted on an acrylic resin). 

Systems such as Pseudomonas lipase dispersed inside Hyflo Supercell (a diatoma-ceous silica of pH 8.5-9) and SP 435 Novozym (Candida antarctica lipase grafted on an acrylic resin) are thermally stable and have optimum activity in the range 80-100 °C. They can therefore be used with conventional or microwave heating if the temperature is strictly controlled. 

The regioselective esterification at position 6 of a-D-glucose and a-D-glucopyrano-sides with fatty acids [81] is readily achieved by use of Novozym 435, in accordance with Scheme 8.56. 

The multipolymer enzymatic resolution of soluble polymer-supported alcohols 42 and 43 was achieved using an immobilised lipase from Candida Antarctica (Novozym 435). The R-alcohol was obtained in enantiomerically pure form (>99% ee) after its cleavage from the poly(ethylene) glycol (PEG) scaffold . The achiral hydantoin- and isoxazoline-substituted dispirocyclobutanoids 47 were produced using both solution and solid-phase synthesis <00JOC3520, OOCC1835>. 

Novozymes supplies the proteases for liquid detergents to the detergent manufacturer as a stable liquid enzyme formulation from which typically less than 2%(w/w) is added to the liquid detergent composition. The limited solubility of boric acid thus prevents Novozymes from supplying the detergent manufacturer with a liquid enzyme formulation with a built-in boric acid stabilization system. 

Further the inhibitor according to the goals should be toxicological safe, be cost-effective compared to boric acid and be compatible with Novozymes enzyme formulations. 

Lone Kierstein Nielsen Novozymes a.s., Liquid Products Development, DK-2800 Bagsvaerd, Denmark... 

Cherry, J. Novozymes, in Proc. Biorefinery 2004, San Francisco, June 8, 2005. 

By screening a variety of lipases in organic solvent for their ability to acylate the racemic hydroxynitrile with succinic anhydride, Novozym 435 was found to yield the best results, affording product in 94-95 % ee at conversions of 47 9 % (Scheme 1.34). After optimization, the reaction was successfully run at 22 kg scale. The immobilized catalyst could be easily isolated by filtration and reused. 

The ester was screened against a panel of enzymes for hydrolysis activity from which only Novozym 435 efficiently hydrolysed the desired (5)-enantiomer." After significant optimization studies using Novozym 435, a process was established where a 100 g slurry of racemic ester in commercial tert-butanol (which is supplied as a mixture containing 12 % water - anhydrous terf-butanol could not be used due to its higher melting point), furnished the desired acid in 43% yield and >99% ee (Scheme 1.36). The reaction was performed at 50 °C as a compromise that gave satisfactory substrate concentration... 

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