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Method for rapidly determining biomass in liquid fermentation liquor

A liquid fermentation and rapid determination technology, applied in the field of biomass determination, can solve the problems of cumbersome operation, high cost, and great difficulty, and achieve the effects of strong correlation, less reagent consumption, and strong operation convenience

Active Publication Date: 2018-10-16
FUZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] There are many methods for measuring biomass, but many of them are cumbersome to operate, and are susceptible to interference from bacterial age, culture conditions, and culture substrates, such as the ergosterol method; in addition, to measure the consumption of carbon and nitrogen sources, Measurements such as respiration rate and dry weight do not always accurately reflect changes in biomass
Although qPCR technology can achieve high-throughput detection and its detection results are relatively accurate, it requires primer design, which is difficult and costly. Urgent problems

Method used

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  • Method for rapidly determining biomass in liquid fermentation liquor
  • Method for rapidly determining biomass in liquid fermentation liquor
  • Method for rapidly determining biomass in liquid fermentation liquor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Monascus (Monascus) Determination of bacterial count

[0035] Insert Monascus spores into the centrifuged fermentation medium without precipitation, and after 3 days of fermentation, centrifuge the fermentation broth at 4500rpm for 10 minutes, discard the supernatant, and obtain the precipitation of Monascus pure bacteria; preparation of the fermentation medium without precipitation by centrifugation: monosodium glutamate 22.33 g, ammonium sulfate 9.72g, anhydrous glucose 42.50g, potassium dihydrogen phosphate 9.00g, manganese sulfate monohydrate 0.21g, magnesium sulfate heptahydrate 2.1g, add water to 1000mL, autoclave at 121℃ for 20 minutes;

[0036] Insert Monascus spores into the fermentation medium with precipitation in centrifugation, centrifuge the fermentation broth at 4500rpm for 10min after 3 days of fermentation, discard the supernatant, and obtain the mixed precipitation of Monascus thallus and medium; prepare the fermentation medium with precipit...

Embodiment 2

[0046] Example 2 Rhizopus oryzae (Rhizopus oryzae) Determination of bacterial count

[0047] The determination steps of the metal ion method are consistent with the operation steps of the amount of Monascus in Example 1. The standard curve between the absorbance value ΔA of Rhizopus oryzae and the mass of pure bacteria is shown in Table 3. The determination comparison between the metal ion method and the glucosamine method is shown in Table 4.

[0048] Table 3 Relevant data of the determination of the amount of Rhizopus oryzae

[0049]

[0050] The standard curve between △A and the mass of pure bacteria is: y=3.7039x-0.0483, R 2 =0.9992.

[0051] Table 4 Comparison of different determination methods

[0052]

[0053] The correlation coefficient of the two determination methods is 0.9835, which means that there is 98.35% certainty that the two determination methods are related.

Embodiment 3

[0054] Example 3 Paecilomyces (Paecilomyces sp.) Determination of bacterial count

[0055] The determination steps of the metal ion method are consistent with the operation steps of the amount of Monascus in Example 1. The standard curve between the absorbance value ΔA of Paecilomyces and the mass of pure bacteria is shown in Table 5. The determination comparison between the metal ion method and the glucosamine method is shown in Table 6.

[0056] Table 5 Relevant data of Paecilomyces determination and standard curve drawing

[0057]

[0058] The standard curve between △A and the mass of pure bacteria is: y=0.1135x-0.0609, R 2 =0.9986.

[0059] Table 6 Comparison of different assay methods

[0060]

[0061] The correlation coefficient of the two determination methods is 0.9825, which means that there is 98.25% certainty that the two determination methods are related.

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Abstract

The invention discloses a method for determining biomass aiming at a micro-porous plate fermentation technology. The method comprises the following steps: cleaning a fermentation liquor precipitate subjected to pore plate culture with normal saline for three times, determining magnesium ions inside thallus based on a microplate reader by virtue of an Eriochrome black T dual-wavelength developing method (Delta A=A560-A680), and reflecting the thallus biomass by using the content of the magnesium ions. According to the method disclosed by the invention, the defect that the biomass of the fermentation liquor is difficultly represented by a dry weight weighing method in the micro-porous plate fermentation process is overcome, and based on elimination of a plurality of interference factors, thedetermined value obtained by the method is close to an actual value, and the method has excellent determination result correlation to other biomass determining methods (such as a glucosamine method)and accords with the growth tendency of bacterial amount in a plate culture count method. In a word, the biomass determining method disclosed by the invention is convenient to operate, rapid and accurate, capable of accurately reflecting the bacterial amount variation tendency in the fermentation process, noncorrosive in reagent and small in dose, and has excellent application value.

Description

technical field [0001] The invention belongs to the field of biomass measurement, in particular to the biomass measurement technology of fungi in liquid fermentation broth. Background technique [0002] There are many methods for measuring biomass, but many of them are cumbersome to operate, and are susceptible to interference from bacterial age, culture conditions, and culture substrates, such as the ergosterol method; in addition, to measure the consumption of carbon and nitrogen sources, Detection methods such as respiration rate and dry weight do not always accurately reflect changes in biomass. Although qPCR technology can achieve high-throughput detection and its detection results are relatively accurate, it requires primer design, which is difficult and costly. Urgent problems. [0003] Since microorganisms can utilize metal ions in the culture environment, the amount of bacteria can be indirectly measured according to the content of metal ions in the bacteria. In ...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/82G01N21/31
CPCG01N21/3151G01N21/78G01N21/82
Inventor 倪莉周康熙冯瀚中何冬萍刘志彬张雯张晨
Owner FUZHOU UNIV
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