A kinetic analysis and optimization of reaction conditions for the enzymatic hydrolysis of chitin using chitinase produced by Trichoderma harzianum NCIM 1185 was carried out. Swollen chitin was used as the substrate for chitinase. The central composite design was followed for this optimization. The required volume ratio of the major reactants for maximum hydrolysis was determined. The pH and temperature optima were found to be 4.75 and 47 °C respectively. K(m) and V(max) for this enzyme were 4.643 kg/m3 and 0.1542 U respectively.

T Panda

Department Of Chemical Engineering

Chitin

CHITINASE

article

controlled study

Enzyme kinetics

Hydrolysis

HYPHOMYCETES

nonhuman

priority journal

Temperature

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IIT Madras

Parameters optimization of chitin hydrolysis by Trichoderma harzianum chitinase under assay conditions

Bioprocess Engineering

Debaryomyces nepalensis was previously isolated, which is capable of producing pectin lyase and pectate lyase using pectin as sole source of carbon. The parameters affecting the activity of these enzymes are categorized into two, viz., chemical (amount of substrate and enzyme) and physical (pH of reaction mixture and temperature of incubation). The effect of these parameters on pectin lyase and pectate lyase was studied using central composite design. The optimal conditions of amount of substrate and enzyme (culture broth) were found to be 545 μl (∼1 g/l pectin) and 123 μl for pectin lyase and 707 μl (∼1.1 g/l polygalacturonic acid) and 96 μl for pectate lyase. The optimum pH and temperature for reaction was found to be 6.4 and 35 °C for pectin lyase and 7.5 and 32 °C for pectate lyase. After optimization, the activity of pectin lyase and pectate lyase was increased by 1.3- and 1.4-fold, respectively. © 2006 Elsevier B.V. All rights reserved.

Biochemical Engineering Journal

Optimization of chemical and physical parameters affecting the activity of pectin lyase and pectate lyase from Debaryomyces nepalensis: A statistical approach

Aqueous two-phase system has already been established to be powerful tool for biomolecule purification, and is increasingly gaining importance as an alternative to conventional extractive fermentation methods. High cost of dextran is the main inhibition in such effort and hence in this paper a much cheaper source of dextran is evaluated as successful phase forming polymer and compared with standard microcrystalline dextran T500. Other polymer used for both the cases was polyethylene glycol (PEG). An exhaustive study has been carried to compare all its properties such as phase composition properties, polymer properties, gel-forming properties, rheological property and partition property. Various fundamental properties of polymer such as critical temperature, specific rotation, intrinsic viscosity, glucose content and dry weight were estimated for this new dextran source. Finally, cost analysis was carried out and tallied for both the dextran fractions. The results of these studies show that significant cost reduction can be achieved without much effect on the process, rather more advantageously, by replacing high cost microcrystalline dextran T500 with relatively cheaper new dextran T40 source. © 2004 Elsevier B.V. All rights reserved.

Evaluation of an alternative source of dextran as a phase forming polymer for aqueous two-phase extractive system

The critical parameters, such as slant age, mycelium age and contact time of mycelium with lytic enzymes which affects the formation of protoplast from the Penicillium griseofulvum mycelium were studied using a novel technique called response surface methodology. The design used to optimize these parameters was full factorial central composite design. The number of protoplasts, viability of protoplasts and griseofulvin yield are the three responses studied in this communication. The individual optimum values of slant age, inoculum age and contact time obtained are 187.1, 30.81, 21.4 h, respectively, for number of protoplast/ml 68.92, 33.01, 9.88 h, respectively, for percentage viability of protoplasts and 230.2, 73.6, 31.2 h, respectively, for griseofulvin. In order to obtain a single optimum value for all the three responses, a multi-response analysis was carried out using a generalized distance approach. The simultaneous optimum values of slant age, inoculum age and contact time are 133.78, 37.1 and 15.97 h, respectively, for all the three responses. © 2003 Elsevier Science B.V. All rights reserved.

Statistical analysis on some critical parameters affecting the formation of protoplasts from the mycelium of Penicillium griseofulvum

Chitinases help degradation of cell walls containing chitin and thus accelerates protoplast formation. It indirectly helps strain improvement and development of new strains which are economically viable for industrial use. Chitinases consist of endochitinase, exochitinase, β-N-acetylglucosaminidases and chitobiases. The endochitinases can be further characterized for better understanding of mechanism of the enzyme reaction. This review covers the recent advances in isolation and characterization of endochitinases and its components, gene encoding sequences and cloning.

Characterization and applications of chitinases from Trichoderma harzianum - A review

The chitinase fermentation process utilizing chitin as the sole carbon source was investigated in a stirred tank bioreactor. Agitator speed of 224 rpm was found to be most suitable for cell growth as well as for chitinase production. Chitinase yield decreased rapidly at higher agitator speed, while decrease in cell yield at higher agitator speed was not rapid. Probably, mass transfer limitation was predominant in the fermentation process at lower agitator speed. Higher agitator speed appears to reduce chitinase production.

Submerged culture production of chitinase by Trichoderma harzianum in stirred tank bioreactors - The influence of agitator speed

Efficient synthesis of a large number of viable protoplasts from Trichoderma longibrachiatum (Trichoderma reesei) mycelium has been found to be influenced by a number of factors. In this case, Trichoderma harzianum NCIM 1185 culture filtrate has been used to prepare protoplasts from Trichoderma longibrachiatum QM 9414. A method has been devised to isolate a large number of viable protoplasts from the mycelium. Detailed analysis of different factors affecting the synthesis of protoplasts from T. longibrachiatum has not yet been reported. The most suitable conditions for protoplasting were as follows: age of the organism in slant, 3 days; mycelium age, 20 h; volume of lytic enzymes, 190 ml; mycelial weight (dry equivalent), 1.66 g; time of contact with lytic enzymes, 2 h; temperature of protoplasting, 30°C; phosphate buffer, 25 mm, pH 6.5; KCl as osmotic stabilizer, 0.7 m. Proper spreading of the mycelium in contact with the lytic enzymes was also found to be necessary. © 1992.

Journal	Data powered by TypesetBioprocess Engineering
Publisher	Data powered by TypesetSpringer Verlag
ISSN	0178515X
Open Access	No