The present disclosure relates to methods, cells, and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interest, where the cells (e.g., Chinese Hamster Ovary (CHO) cells) have reduced or eliminated activity, e.g., expression, of certain host cell proteins, e.g., enzymes including, but not limited to, certain lipases, esterases, and/or hydrolases.

The present application provides anti-PLA2G2D constructs that bind to PLA2G2D (e.g., anti-PLA2G2D antibodies), nucleic acid molecules encoding an amino acid sequence of the anti-PLA2G2D, vectors comprising the nucleic acid molecules, host cells containing the vectors, methods of preparing the anti-PLA2G2D construct, pharmaceutical compositions containing the anti-PLA2G2D construct, and methods of using the anti-PLA2G2D construct or compositions.

The invention discloses a method for improving the extracellular expression level of a foreign protein by means of phospholipase fusion expression. Four proteins, PLA.sub.2, MBP, CBD and SUMO, are used as a fusion tag to construct a fusion gene. Compared with an original protein MOH without any fusion tag, the extracellular expression level and enzymatic activity of all the four fusion proteins are increased to some degree. Among them, the fusion protein using PLA.sub.2 as the fusion tag has the highest expression level, which is 7.4 times higher than that of the original protein. Compared with other fusion tags, PLA.sub.2 has a low molecular weight and the fusion protein having PLA2 as the fusion tag has the highest expression level (up to 12 g.Math.L.sup.1 in a 7 L fermentation tank for high-density fermentation). It is shown that the secretory expression of a foreign protein can be effectively increased by using PLA.sub.2 as a fusion tag.

The present invention relates to a mutant microbial host cell which is deficient in the production of the AgsE protein or in the production of an homologous thereof if compared with a parent microbial host cell which has not been modified and measured under the same conditions. It has been surprisingly found that when the mutant microbial host cell according to the invention is used in a method to produce a compound of interest, for example an enzyme, an improved yield of said compound is obtained if compared to a method in which a parent host cell which has not been modified is used when measured under the same conditions.

A method for producing lysophosphatidylethanolamine 18:1 includes extracting phospholipids including mainly phosphatidylethanolamine from a microorganism of Pseudomonas sp. and treating the extracted phospholipids with phospholipase A2. An alternative method for producing lysophosphatidylethanolamine 18:1 includes treating a microorganism of Pseudomonas sp. directly with phospholipase A2. The lysophosphatidylethanolamine 18:1 can be used as a plant vaccine material for preventing the plants from injuries caused by pathogen infections and/or environmental stresses and accelerating the recovery of plants injured by pathogen infections and/or environmental stresses, and can also be used as a composition for enhancing fruit ripening (color and sweetness) and storage properties, and as it can be used for an application in plant tissues, food products, pharmaceuticals, cosmetics, and agricultural use, it would be very advantageously used in related industries. This invention also provides a method of producing a phosphatidylethanolamine itself from a microorganism of Pseudomonas sp.

The present invention relates to a method for producing an aqueous protein-containing milk or cream fraction, said method comprising using an enzyme having phospholipase C activity.

A bacterial strain secreting fatty acids, the strain inducing fatty acids to be extracellularly secreted by using phospholipase expressed in the periplasmic space of cell. When a method of producing fatty acids by using the bacterial strain secreting fatty acids is used, fatty acids extracellularly secreted are continuously obtained without apoptosis, leading to lower costs and higher production efficiency. Phospholipase, unlike thioesterase, which is a typical fatty-acid degrading enzyme, decomposes phospholipid to produce free fatty acids. Accordingly, by using the substrate specificity of two different phospholipases, a fatty acid having a specific composition can be selectively produced. Unlike in a typical method in which fat is obtained from cells or tissues, fatty acids secreted during cell growth are obtainable by biding to a hydrophobic material without an extraction process using an organic solvent in large quantities. Accordingly, a more economical, environmentally friendly bio-oil production process can be realized.

The present invention relates to a pharmaceutical composition for treating or preventing a disease related to abnormal suppression of regulatory T cell activity comprising a polypeptide comprising a bee venom-PLA2 amino acid sequence exclusive of a leader sequence as an active ingredient. The secretory bee venom-phospholipase A2 of the present invention activates a regulatory T cell and suppress a differentiation of Th1/Th7. Therefore, the present polypeptide can be used as a pharmaceutical composition for treating or preventing a disease related to abnormal suppression of regulatory T cell activity, i.e. autoimmune diseases, allergic diseases, or neurodegenerative diseases.

A method for producing lysophosphatidylethanolamine 18:1 includes treating phospholipids extracted from a microorganisms of Pseudomonas sp. with phospholipase A.sub.2. The lysophosphatidylethanolamine 18:1 can be used as a plant vaccine material for preventing the plants from injuries caused by pathogen infections and/or environmental stresses and accelerating the recovery of plants injured by pathogen infections and/or environmental stresses, and can also be used as a composition for enhancing fruit ripening (color and sweetness) and storage properties, and as it can be used for an application in plant tissues, food products, pharmaceuticals, cosmetics, and agricultural use, it would be very advantageously used in related industries.

A method of producing a par-baked product dough, comprising incorporating into the dough a phospholipase enzyme and a GH8 xylanase.