The present disclosure is generally related to recombinant fungal cells (strains) for use in the commercial scale production of proteins (polypeptides) of interest. Certain embodiments are therefore related to recombinant fungal cells (strains) producing proteins of interest and overexpressing one or more gene(s) encoding a regulatory protein of the disclosure.

This invention discloses a mutated strain of wild type Bacillus mucilaginosus HSCUP-76-8 and a high-density fermentation method thereof. The mutated strain HSCUP-76-8 was assigned an Accession No. CGMCC No. 8481. A two-stage and regulated high-density fermentation method has been established for the production of HSCUP-76-8. In the first stage, parameters are controlled to reduce the viscosity of the fermentation broth and promote the growth of bacteria, thereby allowing the bacteria to reach an amount in the range of 2.010.sup.9 cfu/mL-2.310.sup.9 cfu/ml. In the second stage, nutritional factors and fermentation conditions are controlled to promote sporulation, thereby producing endospores in the range of 1.510.sup.9 cfu/mL-2.010.sup.9 cfu/ml. The fermentation cycle of the two-stage high-density fermentation is 32-48 hours.

The present disclosure provides compositions comprising recombinant bacterial spores. The present disclosure is also directed to vaccine based compositions, which include recombinant bacterial spores that express CTB and a peanut protein(s) on their surfaces. This disclosure also provides methods for administering these compositions as a treatment or prevention of peanut allergy.

A liquid sporulation method is described. A first liquid culture is prepared by adding bacterial cells to a sporulation broth, wherein an optical density (OD.sub.600) of the first liquid culture is in a range of 0.001 to 0.01. The first liquid culture is incubated and an optical density (OD.sub.600) thereof is increased to be in a range of 0.2 to 2.0. Second liquid culture is prepared by adding the incubated first liquid culture to a predetermined amount of additional sporulation broth, wherein an optical density (OD.sub.600) of the second liquid culture is in the range of 0.001 to 0.1, and a ratio of a volume of the second liquid culture to a volume of the first liquid culture is in a range of 10:1 to 150:1. The second liquid culture is incubated so an optical density (OD.sub.600) thereof is increased to be in a range of 1.0 to 4.0.

A liquid sporulation method is described. A first liquid culture is prepared by adding bacterial cells to a sporulation broth, wherein an optical density (OD.sub.600) of the first liquid culture is in a range of 0.001 to 0.01. The first liquid culture is incubated and an optical density (OD.sub.600) thereof is increased to be in a range of 0.2 to 2.0. Second liquid culture is prepared by adding the incubated first liquid culture to a predetermined amount of additional sporulation broth, wherein an optical density (OD.sub.600) of the second liquid culture is in the range of 0.001 to 0.1, and a ratio of a volume of the second liquid culture to a volume of the first liquid culture is in a range of 10:1 to 150:1. The second liquid culture is incubated so an optical density (OD.sub.600) thereof is increased to be in a range of 1.0 to 4.0.

Disclosed herein are therapeutic compositions containing non-pathogenic, germination-competent bacterial spores, for the prevention, control, and treatment of gastrointestinal diseases, disorders and conditions and for general nutritional health.

Described is an isolated strain of the fungus Colonostachys rosea termed BVT Cr-7 useful as a biological control agent for the treatment of plants. The isolated strain, formulations comprising said strain and/or spores derived from said strain may be applied to plants or plant materials in order to improve plant yield, to improve plant growth, or for the treatment or prevention of diseases or pathogens in the plant.

The invention provides a streptomyces graminearus strain, a streptomyces graminearus oil suspension, as well as its preparation method and application thereof. The streptomyces graminearus oil suspension contains streptomyces graminearus conidiospore powder, emulsifier, stabilizer, wetting dispersant, antioxidant, thickener, ultraviolet protectant, preservative and carrier oil. The use of carrier oil as the dispersion medium offers environmental friendliness, excellent permeability, adhesion properties, and resistance to rain washing. Additionally, the carrier oil enhances the spore germination of streptomyces graminearus. Furthermore, it has the advantages of low application cost, slow spore settling speed, easy to shake well when using, stable efficacy, long storage time, suitability for various spraying techniques and a simple preparation method. The spore survival time of the streptomyces graminearus oil suspension is prelonged, with a spore germination rate is as high as 85% after 18 months of storage. It demonstrates effective control against yam, peanut and forest nematode.

The invention provides a streptomyces graminearus strain, a streptomyces graminearus oil suspension, as well as its preparation method and application thereof. The streptomyces graminearus oil suspension contains streptomyces graminearus conidiospore powder, emulsifier, stabilizer, wetting dispersant, antioxidant, thickener, ultraviolet protectant, preservative and carrier oil. The use of carrier oil as the dispersion medium offers environmental friendliness, excellent permeability, adhesion properties, and resistance to rain washing. Additionally, the carrier oil enhances the spore germination of streptomyces graminearus. Furthermore, it has the advantages of low application cost, slow spore settling speed, easy to shake well when using, stable efficacy, long storage time, suitability for various spraying techniques and a simple preparation method. The spore survival time of the streptomyces graminearus oil suspension is prelonged, with a spore germination rate is as high as 85% after 18 months of storage. It demonstrates effective control against yam, peanut and forest nematode.

The present invention provides a method for determining whether or not a test sample contains a phytopathogenic oomycete selectively from two kinds of oomycetes of a phytopathogenic oomycete and a non-phytopathogenic oomycete. The method according to the present invention comprises: (a) putting the test sample on a front surface of a cellulose film; wherein the cellulose film has a thickness of not less than 0.5 micrometers and not more than 3.7 micrometers; (b) leaving the test sample at rest for not less than 4 hours and not more than 8 hours after the step (a); (c) observing a back surface of the film after the step (b); and (d) determining that the test sample contains the phytopathogenic oomycete, if an oomycete is found on the back surface of the film in the step (c).