Novel Composition of Matter and Method for Stimulating the Growth of Beneficial Microorganisms

Patent metadata

Field	Value
Publication number	US-2016376547-A1
Application number	US-201615260435-A
Country	US
Kind code	A1
Filing date	Sep 9, 2016
Priority date	Jan 25, 2008
Publication date	Dec 29, 2016
Grant date	—

Abstract

The invention describes a novel composition of matter obtained from the leaves of green plants, which is useful in promoting the growth of beneficial microorganisms. Specifically, that the invention describes a hydrolysate prepared from plant leaf biomass (leaf biomass hydrolysate or “LBH”) which dramatically stimulates the growth of beneficial microorganisms. Use of LBH as a fermentation substrate can also stimulate rapid production of organic acids and other organic compounds. LBH can be used as a substrate to promote the fermentation-based production of biobased industrial chemicals or biofuels, LBH can be utilized as a prebiotic to promote the growth of beneficial probiotic organisms. In addition, LBH may also be useful in stimulating the fermentation-based production of other products, examples of which include preservatives, antibiotics, antigens, vaccines, amino acids, vitamins, recombinant proteins, bioremediation treatments, and immobilized enzymes.

First claim

1 - 23 . (canceled)

24 - 33 . (canceled)

34 - 64 . (canceled)

65 . A method for producing leaf biomass hydrolysate (LBH) comprising the following steps:
(a) disrupting one or more leaves from tobacco, spinach or alfalfa, or a mixture of these; (b) drying the disrupted leaves to a water activity level of approximately 0.85 or lower; (c) incubating the disrupted leaves in a liquid comprising water, wherein incubating comprises heating the disrupted leaves in the liquid to a temperature of from about 60° C. to about 90° C. for a period of time of ranging from about 15 minutes to about one (1) week; and (d) filtering the liquid after incubation to remove solids, and produce a liquid comprising soluble constituents of LBH.

66 . The method of claim 65 , which further comprises the step of drying the product of step (d) to a powder.

67 . The method of claim 65 , wherein at least one desirable leaf component is removed from the leaves during or after the leaves are disrupted.

68 . The method of claim 65 , wherein in step (c) the disrupted leaves are heated in water to a temperature of from about 75° C. to about 85° C. for a period of time of ranging from about 30 minutes to about 24 hours.

69 . A method for producing a growth medium comprising leaf biomass hydrolysate (LBH) comprising the following steps:
(a) disrupting one or more leaves from tobacco, spinach or alfalfa, or a mixture of these; (b) drying the disrupted leaves to a water activity level of approximately 0.85 or lower; (c) incubating the disrupted leaves in a liquid comprising water, wherein incubating comprises heating the disrupted leaves in the liquid to a temperature of from about 60° C. to about 90° C. for a period of time of ranging from about 15 minutes to about one (1) week; (d) filtering the liquid after incubation to remove solids, and produce a liquid comprising soluble constituents of LBH; and (e) placing the soluble constituents of LBH into a growth medium.

70 . The method of claim 69 , wherein the growth medium is a powder.

71 . The method of claim 69 , wherein the growth medium is a prokaryotic or a eukaryotic growth medium.

72 . The method of claim 69 , wherein at least one desirable leaf component is removed from the leaves during or after the leaves are disrupted.

73 . The method of claim 69 , wherein the filtered liquid of step (d) is dried to a powder prior to being placed into a growth medium.

74 . The method of claim 69 , wherein the growth medium further comprises a protein source and a carbohydrate source.

75 . The method of claim 74 , wherein the protein source is soy and the carbohydrate source is glucose.

76 . The method of claim 69 , wherein the dried, disrupted leaves of step (b) are packaged in filter paper, ground into a powder and packaged, or packaged as disrupted leaves without either filter paper or being ground into a powder, prior to performing the incubating of step (c).

77 . A method to promote the growth of a microorganism selected from the group consisting of E. coli , other Escherichia species, Lactobacillus, Pediococcus, Leuconostoc, Pediococcus, Lactococcus, Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Teragenococcus, Vagococcus, Weisella, Acetobacter, Gluconacetobacter, Clostridium, Aspergillus, Candida, Rhizopus, Actinobacillus, Saccharomyces and Xanthamonas, comprising the steps of preparing LBH by (a) disrupting one or more leaves from tobacco, spinach or alfalfa, or a mixture of these; (b) drying the disrupted leaves to a water activity level of approximately 0.85 or lower; (c) incubating the disrupted leaves in a liquid comprising water, wherein incubating comprises heating the disrupted leaves in the liquid to a temperature of from about 60° C. to about 90° C. for a period of time of ranging from about 15 minutes to about one (1) week; and (d) filtering the liquid after incubation to remove solids, and produce a liquid comprising soluble constituents of LBH; and (e) incubating the microorganism in a growth medium comprising LBH, under conditions that the microorganism is allowed to grow.

78 . The method of claim 77 , wherein the growth medium comprises a component selected from a group consisting of MRS medium, LB medium, TBS medium, yeast extract, soy peptone, casein peptone, meat peptone, and commercially available microbial growth media.

79 . The method of claim 77 , which comprises the further steps of incubating the microorganism under conditions causing the microorganism to produce a fermentation product, and isolating the fermentation product.

80 . The method of claim 79 , wherein the fermentation product is selected from the group consisting of pre-biotics, biofuels, methanol, ethanol, propanol, butanol, alcohol fuels, proteins, recombinant proteins, vitamins, amino acids, organic acids, enzymes, antigens, antibiotics, organic chemicals, bioremediation treatments, preservatives and metabolites.

81 . The method of claim 80 , wherein the organic acid is selected from the group consisting of lactic acid, propionic acid, acetic acid, succinic acid, malic acid, glutamic acid, aspartic acid and 3-hydroxypropionic acid.

82 . The method of claim 79 , wherein the microorganism is a Streptomyces and the fermentation product is an antibiotic.

83 . The method of claim 77 , wherein the microorganism is an E. coli.

84 . The method of claim 77 , wherein the growth medium further comprises a protein source and a carbohydrate source.

85 . The method of claim 84 , wherein the protein source is soy and the carbohydrate source is glucose.

Assignees

• Univ Maryland

Inventors

• Lo Yangming Martin
• Yossa Irene N N
• Belson Neil A

Classifications

• C12N1/22 Primary

  Processes using, or culture media containing, cellulose or hydrolysates thereof · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N1/16

  Yeasts; Culture media therefor · CPC title

• B65B63/08

  for heating or cooling articles or materials to facilitate packaging · CPC title

• B65B55/12

  Sterilising contents prior to, or during, packaging · CPC title