Method for producing organic acid at high production rate through fermentation of intermittent backflow cells

Method for producing organic acid at high production rate through fermentation of intermittent backflow cells

  • CN 102,051,386 B
  • Filed: 10/27/2009
  • Issued: 04/15/2015
  • Est. Priority Date: 10/27/2009
  • Status: Active Grant
First Claim
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1. a method for intermittent type backflow cell high productivity fermentation production of organic acid, is characterized in that, comprise the following steps:

  • Step one, sterilizing;

    steam sterilizing is carried out to the seeding tank produced for organic acid fermentation, fermentor tank, buffer reservoir, filtration unit and connecting tube thereof;

    Step 2, prepare aseptic seed culture medium;

    with the filtration unit through sterilizing, seed culture medium is filtered, obtain aseptic seed culture medium clear liquid, this seed culture medium clear liquid is sent into seeding tank;

    Step 3, prepare highdensity seed liquor;

    the bacterial classification selecting brevibacterium flavum or Corynebacterium glutamicum to be used for organic acid fermentation to produce, the access of selected bacterial classification has been equipped with in the seeding tank of seed culture medium clear liquid, when spawn culture 8h ~ 60h enters the logarithmic phase of its growth curve, the aseptic filtration devices of the seed culture fluid obtained is concentrated, in the seed concentrated solution that filtration unit is retained, the dry weight of biomass reaches 30g/L, and this seed concentrated solution is high-density seed liquor;

    Step 4, prepare aseptic fermention medium;

    fermention medium that organic acid fermentation produces will be used for by after aseptic filtration devices, obtain aseptic fermention medium clear liquid, a part for this fermention medium clear liquid is sent into fermentor tank, and another part is deposited in aseptic buffer reservoir for subsequent use;

    Step 5, intermittent type backflow cell method fermentation production of organic acid;

    the high-density seed liquor prepared by step 3 is inoculated in fermentor tank, be conducive to cultivating under selected bacterial classification synthesis organic acid fermentation condition, described fermentation condition is set to;

    when selecting brevibacterium flavum to be used for the bacterial classification of organic acid fermentation production, pass into ammonia by pH value modulation 6.5, blowing air amount 1.2L/min, mixing speed 200rpm, 32 DEG C of constant temperature;

    And when selecting Corynebacterium glutamicum to be used for the bacterial classification of organic acid fermentation production, stream adds the Na of 6mol/L 2cO 3solution adjusts pH to 6.8, air flow 1.5L/min, mixing speed 200rpm, 32 DEG C of constant temperature;

    Monitor the remaining sugar concentration of fermentation cylinder for fermentation liquid in fermenting process, when fermented liquid remaining sugar concentration is down to 6.3g/L, carry out the operation of following (1) and (2) simultaneously;

    (1) fermented liquid is discharged in fermentor tank, the long-pending change of fermentation cylinder for fermentation liquid is monitored in the process that fermented liquid is discharged, discharge is stopped when the fermented liquid in fermentor tank has discharged 100% of its cumulative volume, fresh aseptic fermention medium clear liquid is supplemented immediately in fermentor tank, the fresh fermention medium volume supplemented is not more than the fermentating liquid volume of discharge, so the sugared concentration in fermentor tank is greater than concentration when fermented liquid is just discharged in fermentor tank;

    (2) fermented liquid of discharge is sent into aseptic filtration devices, to be membrane pore size be this filtration unit is no more than the micro-filtration of 0.22 μ

    m, the fermentation clear liquid leached, for extracting organic acid, sends back to fermentor tank again continue to use as production bacterium using being filtered somatic cells concentrated solution that device retains;

    Remaining sugar concentration scope when the remaining sugar concentration in fermentor tank is down to first time discharge fermented liquid again, and compared with first time discharges before fermented liquid, when declining does not appear in organic acid productivity, again carry out the operation of above (1) and (2) simultaneously;

    Remaining sugar concentration scope when the remaining sugar concentration in fermentor tank is down to first time discharge fermented liquid again, and compared with first time discharges before fermented liquid, organic acid productivity occur decline and when being no more than 10%, in fermentor tank, supplement the high-density seed liquor prepared by method described in abovementioned steps three immediately, and again carry out the operation of above (1) and (2) simultaneously;

    Remaining sugar concentration scope when the remaining sugar concentration in fermentor tank is down to first time discharge fermented liquid again, and compared with first time discharges before fermented liquid, organic acid productivity occur decline and more than 10% time, fermented liquid in fermentor tank is all discharged, aseptic filtration devices sent into by the fermented liquid of discharging, the fermentation clear liquid leached is for extracting organic acid, the somatic cells concentrated solution retained no longer sends back to fermentor tank, but pass into disuse, so far namely complete one and take turns complete intermittent type backflow cell high productivity fermentation production of organic acid process;

    Step 6, new round intermittent type backflow cell method fermentation production of organic acid;

    monitor whole production system and whether occur microbiological contamination, repeating above-mentioned steps one to five when there is microbiological contamination, continuing the intermittent type backflow cell high productivity fermentation production of organic acid process of a new round;

    When not there is microbiological contamination, repeating above-mentioned steps two to five, continuing the intermittent type backflow cell high productivity fermentation production of organic acid process of a new round.

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