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Bacterial fermentation methods and compositions

A technology for compositions and fermented products, applied in the directions of bacteria, botanical equipment and methods, and methods of using microorganisms, etc., can solve problems such as equipment cannot be constructed, and achieve the effect of shortening production time

Inactive Publication Date: 2016-04-13
NEWLEAF SYMBIOTICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Such equipment may not be constructed or operated in a commercially viable

Method used

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  • Bacterial fermentation methods and compositions
  • Bacterial fermentation methods and compositions
  • Bacterial fermentation methods and compositions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1. Growth of PPFM bacteria on solid agar plates

[0093] Various standard media were tested for the growth of PPFM bacteria on solid agar plate media.

[0094] One medium used was Ammonium Mineral Salts (AMS) medium (Whittenbury et al. 1970). Each liter of AMS medium contains 700 mg of dipotassium hydrogen phosphate anhydrous, 540 mg of potassium dihydrogen phosphate, 1 g of magnesium sulfate heptahydrate, 500 mg of anhydrous ammonium chloride, 200 mg of calcium chloride dihydrate, 4 mg of sulfuric acid heptahydrate Iron, 100 mcg zinc sulfate heptahydrate, 30 mcg manganese chloride tetrahydrate, 300 mcg anhydrous boric acid, 200 mcg cobalt chloride hexahydrate, 10 mcg copper chloride dihydrate, 20 mcg nickel chloride hexahydrate, and 60 mcg dihydrate Sodium molybdate hydrate.

[0095] AMS medium was prepared from the four stock solutions listed below

[0096] Stock solution I: For 1 L at 50x concentration

[0097] Dipotassium Phosphate, Anhydrous 35g

[009...

Embodiment 2

[0122] Example 2. Growth of PPFM bacteria in clarified single-phase liquid medium

[0123] For those four solid agar plate media found in Example 1 that supported the fastest and most abundant growth of the PPFM bacterium M. extrusive, the corresponding liquid forms (ie, no agar added) were prepared and tested. The four liquid media prepared as described in Example 1 (with the only exception that they did not contain any agar) were water-clear liquids with all components in solution.

[0124] To a flask containing 100 ml of these four liquid media, add an inoculum of the PPFM bacteria Methylobacter extorts to yield approximately 1 x 10 5 Initial titer of colony forming units (CFU) / ml. The flasks were placed in a rotary shaker incubator setup and grown at 30°C at 250 rpm for 5 days. At the end of the 5-day incubation, the titers of PPFM bacteria in the flasks were determined. The result is:

[0125]

[0126]

[0127] The surprising aspect of these results is that in t...

Embodiment 3

[0128] Example 3. Growth of PPFM bacteria in biphasic medium containing insoluble salt crystals

[0129] To prepare the biphasic medium, the liquid AMS+glycerol and peptone medium was turbid (ie provided with solid matter) by the intentional formation of insoluble crystals of magnesium phosphate and / or calcium phosphate. In order to intentionally form insoluble crystals in the medium, the preparation method described in Example 1 was modified as follows. All components except the trace metal stock solution were mixed together prior to autoclaving. That is, to 940 ml of distilled water were added 20 ml of each of stock solutions I, II and III and 10 grams of glycerol and 10 grams of peptone. After autoclaving, the medium was completed by adding 1 ml of filter sterilized trace metal stock solution. Autoclaving the components of stock solutions I, II and III that were mixed together prior to autoclaving resulted in the formation of insoluble salt crystals presumably primarily m...

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Abstract

The present invention provides methods for the cultivation of the Methylobacterium genus of bacteria. In particular the method provides methods for the efficient and inexpensive cultivation of these bacteria. Additionally, the invention provides methods for the utilization of these bacterial cultures to improve plant agriculture.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This application claims the benefit of US Patent Application Serial No. 61 / 829,987, filed May 31, 2013, the entire disclosure of which is incorporated herein by reference. Background of the Invention [0003] One-carbon organic compounds, such as methane and methanol, are widely present in nature, and bacteria classified as methanotrophs and methylotrophs are used as carbon sources. Methanogenic bacteria include species in the following genera: Methylobacter, Methylomonas, Methylomicrobium, Methylococcus, Methylococcus The genera Methylosinus, Methylocystis, Methylosphaera, Methylocaldum, and Methylocella (Lidstrom, 2006). Methanogens have methane monooxygenase, which will be derived from O 2 The oxygen atoms are incorporated into methane to form methanol. All methanotrophs are obligate one-carbon users and cannot utilize compounds containing carbon-carbon bonds. On the other hand, methylotrophs can also utilize more ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/30A01N63/20
CPCC12N1/20A01N63/20Y02E50/30
Inventor 格雷格·博戈西安
Owner NEWLEAF SYMBIOTICS INC
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