METHODS AND COMPOSITIONS FOR CULTURING HEMOGLOBIN-DEPENDENT BACTERIA
Provided herein are compositions and methods related to culturing bacteria (such as hemoglobin-dependent bacteria) with a heme-containing polypeptide (such as a heme-containing polypeptide that is not sourced from an animal and/or can be recombinantly produced).
1 . A method of culturing hemoglobin-dependent bacteria, the method comprising incubating the hemoglobin-dependent bacteria in a growth medium that comprises a heme-containing polypeptide, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.
2 . The method of claim 1 , wherein the heme-containing polypeptide is a hemoglobin.
3 . (canceled)
4 . The method of claim 1 , wherein the heme-containing polypeptide is a leghemoglobin.
5 . The method of claim 1 , wherein the heme-containing polypeptide is a myoglobin.
6 . The method of claim 1 , wherein the heme-containing polypeptide is a piscine polypeptide.
7 - 8 . (canceled)
9 . The method of claim 1 , wherein the heme-containing polypeptide is an avian polypeptide.
10 - 11 . (canceled)
12 . The method of claim 1 , wherein the non-animal-derived polypeptide is purified from a plant, bacteria, cyanobacteria, fungus, algae, grain, legume, and/or protozoa.
13 - 18 . (canceled)
19 . The method of claim 1 , wherein the heme-containing polypeptide comprises an amino acid sequence with at least 70% identity to an amino acid sequence in any one of SEQ ID NOs: 1-29, 31, 33, 35, 37, 39, 41, 43, 114, or a combination thereof.
20 . (canceled)
21 . The method of claim 1 , wherein the heme-containing polypeptide comprises an amino acid sequence with at least 70% identity to the amino acid sequence of SEQ ID NO: 4.
22 - 23 . (canceled)
24 . The method of claim 1 , wherein the heme-containing polypeptide comprises an amino acid sequence with at least 70% identity to the amino acid sequence of SEQ ID NO: 114 or UniProtKB—P02236 (LGB2_SOYBN).
25 - 33 . (canceled)
34 . The method of claim 12 , wherein the heme-containing polypeptide is a soy leghemoglobin recombinantly expressed in Pichia pastoris.
35 . The method of claim 1 , wherein the hemoglobin-dependent bacteria are bacteria of the genus Actinomyces, Alistipes, Anaerobutyricum, Bacillus, Bacteroides, Cloacibacillus, Clostridium, Collinsella, Cutibacterium, Eisenbergiella, Erysipelotrichaceae, Eubacterium/Mogibacterium, Faecalibacterium, Fournierella, Fusobacterium, Megasphaera, Parabacteroides, Peptoniphilus, Peptostreptococcus, Porphyromonas, Prevotella, Propionibacterium, Rarimicrobium, Shuttleworthia, Turicibacter , or Veillonella.
36 . The method of claim 1 , wherein the hemoglobin-dependent bacteria are of the genus Prevotella.
37 - 38 . (canceled)
39 . The method of claim 36 , wherein the Prevotella comprise at least 99% genomic, 16S and/or CRISPR sequence identity to the nucleotide sequence of the Prevotella Strain B 50329 (NRRL accession number B 50329).
40 . (canceled)
41 . The method of claim 36 , wherein the Prevotella comprise at least 99% genomic, 16S and/or CRISPR sequence identity to the nucleotide sequence of the Prevotella Strain C (PTA-126140).
42 . (canceled)
43 . The method of claim 36 , wherein the hemoglobin-dependent bacteria are a strain of Prevotella bacteria comprising one or more proteins listed in Table 1.
44 . The method of claim 36 , wherein the hemoglobin-dependent bacteria are from a strain of Prevotella substantially free of a protein listed in Table 2.
45 - 48 . (canceled)
49 . A method of culturing hemoglobin-dependent bacteria, the method comprising (a) adding a heme-containing polypeptide and hemoglobin-dependent bacteria to a growth medium; and (b) incubating the hemoglobin-dependent bacteria in the growth medium, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.
50 . A bioreactor comprising hemoglobin-dependent bacteria in a growth medium comprising a heme-containing polypeptide, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.
51 - 52 . (canceled)
53 . A composition comprising
a) hemoglobin-dependent bacteria, and
b) a growth medium comprising a heme-containing polypeptide, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.
54 . A growth medium for use in culturing hemoglobin-dependent bacteria, the growth medium comprising a heme-containing polypeptide, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.
55 . A heme-containing polypeptide for use as a supplement in a growth medium for facilitating the growth of hemoglobin-dependent bacteria, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.
56 . A bacterial composition comprising
(a) hemoglobin-dependent bacteria, and
(b) a heme-containing polypeptide, wherein the heme-containing polypeptide is:
(i) a piscine polypeptide;
(ii) an avian polypeptide; or
(iii) a non-animal-derived polypeptide.