Acceleration of hyaluronidase production in the course of batch cultivation of Clostridium perfringens can be achieved with bacteriolytic enzymes

Abstract

As Clostridium perfringens hyaluronidase has cell‐bound enzyme features, an enzymatic approach has been designed to facilitate the release of hyaluronidase into culture through increasing the clostridial cell wall permeability. As a result of the application of lytic peptidase from Actinomyces rutgersensis,β‐N‐acetylglucosaminidase and β‐N‐acetylmuramidase (both from Bacillus subtilis) commercially available preparations at the end of the producer's exponential growth phase, a 5·3‐, 4·8‐ and 4·0‐fold acceleration, respectively (but no enhancement), of hyaluronidase production in the course of batch cultivation of Cl. perfringens has been achieved. This also resulted in an approximately 10‐fold reduction in undesirable side lecithinase activity irrespective of the bacteriolytic enzyme preparation used.

Hyaluronidase

“…This enzyme hydrolyzes HA by splitting the β 1,4‐glucosaminidic bond between C1 of the glucosamine moiety and C4 of the glucuronic acid (Figure 2). It also serves a number of natural biological functions, including human fertilization and the enhancement of bacterial pathogenicity by allowing greater access and mobility through human body tissues 7,8 …”

“…It also serves a number of natural biological functions, including human fertilization and the enhancement of bacterial pathogenicity by allowing greater access and mobility through human body tissues. 7,8 This article reviewing hyaluronidase will be broken down into five sections: various commercial hyaluronidase formulations; adverse effects, skin testing, and precautions; pharmacokinetics; FDA-approved uses of hyaluronidase; and off-label uses of hyaluronidase in dermatology.…”

Enzymatic lysis of microbial cells

“…For instance, Yang et al (2000) used a temperature-sensitive lytic system for efficient recovery of recombinant proteins from Escherichia coli, and Zukaite and Biziulevicius (2000) applied lytic enzymes to accelerate the production of hyaluronidase by recombinant Clostridium perfringens in the course of batch cultivation.…”

“…In consequence, besides the use of Triton X-100 or EDTA, several other strategies have been developed to expand the uses of lysozyme to Gram-negative bacteria. These include: denaturation of lysozyme (Touch et al 2003); modification of lysozyme by covalent attachment of polysaccharides (Aminlari et al 2005) or fatty acids (Ibrahim et al 1991); attachment of hydrophobic peptides to the C-terminal Kerr and Wellnitz (2003), Donovan et al (2005) Antimicrobial for medical and food applications Masschalck and Michiels (2003), Loeffler et al (2001), Fischetti (2003), Sava (1996) Release of recombinant proteins Zhang et al (1999), Zukaite and Biziulevicius, (2000), de Ruyter et al (1997) Yeast-lysing enzymes Preparation of protoplasts, cell fusion, and transformation of yeast Kitamura (1982) Production of intracellular enzymes Zomer et al (1987) Pre-treatment to increase yeast digestibility Kobayashi et al (1982) (Ibrahim et al 1994) or cell permeabilization by high hydrostatic pressure (Masschalck et al 2001). Extensive hydrolysis of peptidoglycan by lysozyme results in cell lysis and death in a hypo osmotic environment.…”

Determination of the efficient enzyme concentration for lytic digestion of vegetative cells but not spores in Schizosaccharomyces pombe

“…For instance, Yang et al (2000) used a temperature-sensitive lytic system for efficient recovery of recombinant proteins from Escherichia coli. Moreover, Žukaite and Biziulevičius (2000) applied lytic enzymes to enhance the production of hyaluronidase by recombinant Clostridium perfringens in the course of batch cultivation. Digestion of yeast and fungal cell walls is necessary for many experimental procedures.…”