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.

T Panda

Department Of Chemical Engineering

J. Anjani Kumari

BIOREACTORS--EFFICIENCY

CHEMICAL REACTIONS--HYDROLYSIS

FUNGAL CELL CULTURES

FUNGAL PROTOPLAST PRODUCTION

LYTIC PROTOPLASTING ENZYMES

OPTIMAL PROTOPLASTING CONDITIONS

STIRRED-TANK BIOREACTORS

Cell culture

hydrolase

article

culture medium

nonhuman

PROTOPLAST

TRICHODERMA

TRICHODERMA REESEI

HYPOCREA JECORINA

HYPOCREA LIXII

LONGIBRACHIATUM

TRICHODERMA LONGIBRACHIATUM

IIT Madras is a public technical and research university located in Chennai, Tamil Nadu. Founded in 1959, it is recognised as an Institute of National Importance.

IIT Madras has been ranked as the top engineering institute in India for four years in a row by the National Institutional Ranking Framework of the MHRD

It currently offers undergraduate, postgraduate and research degrees across 16 disciplines in Engineering, Sciences, Humanities and Management. About 596 faculty belonging to science and engineering departments and centres of the Institute are engaged in teaching, research and industrial consultancy.

IIT Madras

Studies on critical analysis of factors influencing improved production of protoplasts from Trichoderma reesei mycelium

Enzyme and Microbial Technology

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.

Biochemical Engineering Journal

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.

Bioprocess Engineering

Characterization and applications of chitinases from Trichoderma harzianum - A review

Production of microbial chitinases have received increased attention in recent years due to its potential application in various fields. There has been an all around effort to increase the production of chitinases by using genetically engineered organisms and by incorporating modern fermentation techniques. This review presents a brief outline on the production of microbial chitinases which includes information on random screening and selection of chitinolytic organisms and the rationale behind the use of genetically engineered organisms. The problems and perspectives involved in large scale production of chitinases is discussed with special reference to bioreactor studies and new fermentation methods. The present limitations in the understanding of the chitinase fermentation process is also discussed and the scope for future investigation is outlined in this communication.

Production of microbial chitinases - A revisit

A novel optimization procedure has been developed to determine the best combination of variables in large scale fermentation systems. The self- directing optimization or the rotating simplex method of optimization was employed to determine the best suitable combination of parameters, pH (controlled), aeration rate and agitation rate for maximal production of chitinase by Trichoderma harzianum in batch culture. Extracellular enzyme activity increased from an initial value of 0.109 U under unoptimized conditions to a final value of 0.384 U after optimization. A controlled pH of 4.9, an aeration rate of 1.51 l-1 per min, and an agitation rate of 224 rpm was found to be the best combination.

Process Biochemistry

Self-directing optimization of parameters for extracellular chitinase production by Trichoderma harzianum in batch mode

A range of chitinase genes from microorganisms have been cloned and the potential uses of these genetically manipulated organisms are being investigated by various researchers. Fungi and yeast are better producers of chitinase than bacteria. Since fungi grow at a slower rate, there have been efforts to clone the fungal chitinase genes into fast-growing bacteria. This review gives a brief survey of recent progress in the regulation and cloning of microbial chitinase genes. Emphasis is placed on the post-translational modification and localization of the recombinant protein in the host. Various amino acid domains are present in this protein. The mode of catalytic activity of the recombinant protein in comparison to the wild-type protein is discussed in the available literature. The different mechanisms involved in the regulation of chitinase genes from various microorganisms is discussed by the researchers. The scope of future research and conclusions yet to be obtained in this particular area are also outlined in this review.

Applied Microbiology and Biotechnology

Regulation and cloning of microbial chitinase genes

Numerical Methods for Engineers and Scientists

* = = =

Encyclopedia of Genetics, Genomics, Proteomics and Informatics

Bradford Method (Anal Biochem 72:248)

Canadian Journal of Microbiology

Trichoderma protoplast fusion: a tool for improving biocontrol agents

Experimental Mycology

Limited vegetative compatibility following intra- and interspecific protoplast fusion inTrichoderma

Agricultural and Biological Chemistry

Intergeneric hybridization between Aspergillus niger and Trichoderma viride by protoplast fusion.

Journal	Enzyme and Microbial Technology
ISSN	01410229
Open Access	No