The production of microbial polysaccharides has recently gained much interest because of their potential biotechnological applications. Several pathogenic bacteria are known to produce capsular polysaccharides, which provide a protection barrier towards harsh environmental conditions, and towards host defences in case of invasive infections. These capsules are often composed of glycosaminoglycan-like polymers. Glycosaminoglycans are essential structural components of the mammalian extracellular matrix and they have several applications in the medical, veterinary, pharmaceutical and cosmetic field because of their peculiar properties. Most of the commercially available glycosaminoglycans have so far been extracted from animal sources, and therefore the structural similarity of microbial capsular polysaccharides to these biomolecules makes these bacteria ideal candidates as non-animal sources of glycosaminoglycan-derived products. One example is hyaluronic acid which was formerly extracted from hen crests, but is nowadays produced via Streptococci fermentations. On the other hand, no large scale biotechnological production processes for heparin and chondrotin sulfate have been developed. The larger demand of these biopolymers compared to hyaluronic acid (tons vs kilograms), due to the higher titre in the final product (grams vs milligrams/dose), and the scarce scientific effort have hampered the successful development of fermentative processes. In this paper we present an overview of the diverse applications and production methods of chondroitin reported so far in literature with a specific focus on novel microbial biotechnological approaches.

Production of chondroitin sulfate and chondroitin

Several commercial uses and potential novel applications have recently been described for chondroitin sulfate (CS). However, the currently applied animal extractive procedure has a high environmental impact, which may become more profound especially in relation to the forecasted expansion of the CS market for applications as a food supplement, pharmaceutical ingredient, and biopolymer in materials for regenerative medicine. This issue, together with religious and consumer concerns, has prompted the good manufacturing practice (GMP) of chondroitin and CS. This is achievable by combining the design of metabolically engineered microorganisms and tailor-made fermentation processes with semi-synthetic or enzyme-based approaches. The final target is to obtain molecules with specific sulfation patterns that resemble those occurring in natural products and improve the sulfation motif or introduce specific substitutions, such as fucosylation, to tune the biological function. The frontier that is currently triggering attention is related to evaluating the bioactivity of unsulfated chondroitin. Due to recent advancements in the field, a brief survey of the most recent patent and research literature is discussed here.

Microbial production and metabolic engineering of chondroitin and chondroitin sulfate

The invention relates to a screening method for producing chondroitin sulfate bacillus subtilis and application of a bacterial strain fermentation method in the production of chondroitin sulfate, belonging to the technical field of biological engineering. The bacterial strain disclosed by the invention is bacillus subtilis which is obtained by the following steps of: diluting and coating fermented soya beans subjected to a boiling water bath, dyeing and carrying out microscopic examination on an obtained soporiferous strain, then carrying out primary fermentation and shaking culture one by one on screened bacterial strains, adding chondroitin sulfate standard substance of sharks in a formation liquor to be used as an internal standard, carrying out qualitative analysis and screening by adopting a high performance liquid chromatography, and carrying out morphological, physiological and biochemistric and molecular biological identification on the screened bacterial strains to obtain thebacillus subtilis for producing the chondroitin sulfate. Meanwhile, the invention discloses a method for producing the chondroitin sulfate by using the bacterial strain; and the method is used for fermenting the chondroitin sulfate for 24h, wherein the yield of the chondroitin sulfate is 177mg/L.

Screening method for producing chondroitin sulfate bacterial strain and application of bacterial strain fermentation method in production of chondroitin sulfate

The present invention concerns a shark-like chondroitin sulphate and a process for the preparation thereof. In particular, the present invention relates to a shark¬ like chondroitin sulphate, showing a very low amount of 4-sulphate, a high charge density and a biological activity comparable to natural chondroitin sulphates; the invention also relates to a process for the preparation of said shark-like chondroitin sulphate affording substantially higher productivities and better reproducibility of product quality. The shark-like chondroitin sulphate of the invention shows a high molecular mass and charge density; its in vitro biological and anti-inflammatory effectiveness, comparable to the ones of natural products make this polysaccharide potentially useful as a drug in pharmaceutical preparations and nutraceuticals.

Shark-like chondroitin sulphate and process for the preparation thereof

Disclosed is a low-molecular-weight (1000-5000 daltons) chondroitin sulphate (CS) produced by chemical sulphation and subsequent depolymerisation of a non-sulphated chondroitin backbone obtained with biotechnology techniques. The CS described is substantially monosulphated, mainly at the 6-position, with very little sulphation at the 4-position, and with a mono/disulphated disaccharide ratio and charge density similar to those of natural CS. Said biotechnological chondroitin 6-sulphate (C6S) is useful in the treatment and prevention of osteoarthritis and in acute and chronic inflammatory processes.

Low-Molecular-Weight Biotechnological Chondroitin 6-Sulphate for Prevention of Osteoarthritis

Described is chondroitin sulfate obtained from microbial sources, and related compositions and methods.

Microbial-derived chondroitin sulfate

L'invention concerne une enzyme digestant les dipeptides qui agit sur un dipeptide et le digere en acides amines. L'invention concerne egalement un nouveau procede de dosage qui rend possible de doser de maniere rapide, pratique et precise une proteine glyquee, un peptide glyque ou un dipeptide glyque en traitant un dipeptide glyque avec l'enzyme decrite precedemment pour former un acide amine glyque et un acide amine, puis en identifiant l'acide amine. L'invention concerne notamment une enzyme digestant les dipeptides qui agit sur un dipeptide et forme des acides amines. L'enzyme digerant les dipeptides est telle que decrite precedemment, le dipeptide etant un dipeptide glyque et un acide amine glyque et un acide amine etant ainsi formes. L'invention concerne egalement un procede de formation d'un acide amine glyque et d'un acide amine, ledit procede comprenant le traitement d'un echantillon contenant une proteine glyquee ou un peptide glyque avec une protease et une enzyme digerant les dipeptides. L'invention concerne egalement un procede de dosage d'une proteine glyquee, d'un peptide glyque ou d'un dipeptide glyque, ledit procede comprenant la quantification de l'acide amine forme par le procede decrit precedemment, par ajout d'une substance capable de digerer l'acide amine.

Nouvelle enzyme digestant les dipeptides, son procédé de préparation, procédé de dosage des protéines glyquées utilisant l'enzyme digestant les dipeptides et composition de réactif destinée à être utilisée dans ce procédé

Kyouichi Nishio

Papers

Microbial-derived chondroitin sulfate

Nouvelle enzyme digestant les dipeptides, son procédé de préparation, procédé de dosage des protéines glyquées utilisant l'enzyme digestant les dipeptides et composition de réactif destinée à être utilisée dans ce procédé