Microorganisms and methods for reducing bacterial contamination
Inventors
Wong, Steve Good-Sung • Jansen van Vuuren, Hendrik Jurgens
Assignees
Creatus Biosciences Inc • Melio Peptide Systems Inc
Publication Number
US-11788053-B2
Publication Date
2023-10-17
Expiration Date
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Abstract
Provided herein are compositions and methods for reducing bacterial contamination during cell culture. Such compositions and methods utilize engineered peptides or recombinant cells capable of secreting such peptides into culture medium. Also provided are methods of using the engineered peptides for inhibiting bacterial growth during culturing of cells.
Core Innovation
Provided herein are compositions and methods for reducing bacterial contamination during cell culture. Such compositions and methods utilize engineered peptides or recombinant cells capable of secreting such peptides into culture medium. Also provided are methods of using the engineered peptides for inhibiting bacterial growth during culturing of cells.
Commercial fermentation of yeast cells, such as fermentation to produce ethanol, occur in non-sterile environments and frequently suffer from bacterial contamination. Lactic acid bacteria, which are common contaminants, can compete with the yeast cells for nutrients and inhibit yeast cell growth by producing lactic acid and acetic acid, and this production of lactic acid and acetic acid can lead to reduced ethanol yields, "stuck" fermentations, and economic loss.
Small molecule antibiotics, such as virginiamycin, can inhibit bacterial growth and prevent ethanol yield loss, but it is desirable to reduce antibiotic use due to rising concerns of antibiotic resistance in bacteria and demand for antibiotic-free distillers grains. Pediocin PA-1 does not have antibacterial activity against all lactic acid bacteria, including those that are known to contaminate commercial fermentations of yeast; thus, there exists a need to identify alternatives to antibiotics and other peptides that can broaden the spectrum of antibacterial activity against lactic acid bacteria, and the compositions and methods described herein satisfy this need and provide related advantages.
Claims Coverage
The provided claims include one independent claim directed to an engineered persulcatusin, yielding one principal inventive feature.
Engineered persulcatusin with specific amino acid substitutions
A variant of the amino acid sequence of SEQ ID NO: 1, wherein the variant amino acid sequence comprises one or more amino acid substitutions at positions selected from F6, N7, G9, R13, H14, R16, 118, R20, R21, A26, L28, F29, and R38 of SEQ ID NO: 1.
The independent claim is directed to an engineered persulcatusin variant of SEQ ID NO: 1 with one or more amino acid substitutions at specified positions.
Stated Advantages
Reducing bacterial contamination during cell culture.
Inhibiting bacterial growth during culturing of cells via engineered peptides or recombinant cells that secrete such peptides into culture medium.
Engineered peptides with improved antibacterial activity against lactic acid bacteria that are known contaminants of cell cultures.
Providing alternatives to antibiotics and broadening the spectrum of antibacterial activity against lactic acid bacteria, addressing concerns about antibiotic resistance and demand for antibiotic-free distillers grains.
Enabling protection of yeast fermentations and use in production of bioderived compounds (e.g., ethanol) by inhibiting contaminating lactic acid bacteria.
Documented Applications
Inhibiting bacterial growth in a yeast culture by culturing a recombinant yeast described herein or culturing the yeast in the presence of an engineered persulcatusin or engineered pediocin PA-1, including culturing by fermentation.
Co-culturing yeast in the presence of a recombinant yeast that expresses and secretes an engineered peptide to inhibit contaminating lactic acid bacteria.
Culture media comprising engineered persulcatusin or engineered pediocin PA-1, alone or in combination (and optionally containing a bioderived compound), for use in environments susceptible to lactic acid bacteria contamination.
Use of an engineered persulcatusin, an engineered pediocin PA-1, or a recombinant yeast described herein in the production of bioderived compounds selected from ethanol, xylitol, n-butanol, isobutanol, isopropanol, arabitol, ethyl acetate, phenyl-ethyl alcohol, 2-methyl-butanol, 3-methyl-butanol and a combination of carbonyl compounds, alcohols, acetals, acids, esters and nitrogen compounds found in wine, beer, or whiskey.
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