CN102605015A - L-alanine production method - Google Patents

Abstract

The invention discloses an L-alanine production method comprising the following steps: activating passage pseudomonas dacunhae on a sterile agarslantculture-medium; inoculating and culturing the activated pseudomonas dacunhae in a sterile shake-flask seed liquid medium; when the net gain of OD ranges from 0.5 to 0.6, and the pH value ranges from 8.0 to 9.0 and is constant, adding aspartate in a flow way, and controlling the lowest pH value at 6.0; and when the pH value is 6.0 and is constant, finishing fermentation. The shake-flask seed liquid medium comprises 1% of sodium glutamate, 0.8% of peptone, 0.5% of sodium chloride, 0.1% of magnesium sulfate, 0.05% of monopotassium phosphate and 0.55% of corn powder; and the agarslantculture-medium comprises 1% of sodium glutamate, 0.8% of peptone, 0.5% of sodium chloride, 0.1% of magnesium sulfate, 0.05% of monopotassium phosphate, 0.5% of yeast extract, 0.5% of beef extract and 2% of agar. According to the method, the yield and purity of produced L-alanine are high.

Description

Produce the method for L-L-Ala

Technical field

The present invention relates to a kind of method of the L-of production L-Ala.

Background technology

The L-L-Ala (L-Alanine) also claim α-An Jibingsuan; Be one of amino acid that life entity is interior with carbohydrate metabolism is closely related, demand is more; In industries such as medicine, food and chemical industry, be used widely; And become the another important amino acid industry after L-glutamic acid, Methionin, it is very necessary greatly developing production.Present L-L-Ala working method is for being main raw materials for production with fumaric acid, and whole cell enzyme catalysis biological respinse method is produced the L-L-Ala.

Summary of the invention

The method that the purpose of this invention is to provide a kind of L-of production L-Ala.

The method of production L-L-Ala provided by the present invention comprises the steps:

The activation moral A Kun that goes down to posterity breathes out pseudomonas on aseptic slant medium; Moral A Kun Kazakhstan pseudomonas after the activation is inoculated in the aseptic shake-flask seed liquid culture medium to be cultivated; When OD has a net increase of 0.5～0.6, the pH value be 8.0～9.0 continuous when constant stream add aspartic acid, control pH value is minimum to be 6.0; When the pH value is 6.0, accomplish fermentation when constant continuously;

Said shake-flask seed liquid culture medium is Sodium Glutamate 1%, peptone 0.8%, sodium-chlor 0.5%, sal epsom 0.1%, potassium primary phosphate 0.05%, corn dry powder 0.55%.

Said slant medium is Sodium Glutamate 1%, peptone 0.8%, sodium-chlor 0.5%, sal epsom 0.1%, potassium primary phosphate 0.05%, yeast extract paste 0.5%, Carnis Bovis seu Bubali cream 0.5%, agar 2%.

The method of production L-L-Ala of the present invention, the output of L-L-Ala and purity are high.

Embodiment

Following percentage composition is the quality percentage composition like no specified otherwise.

Embodiment

Shake-flask seed liquid culture medium: Sodium Glutamate 1%, peptone 0.8%, sodium-chlor 0.5%, sal epsom 0.1%, potassium primary phosphate 0.05%, corn dry powder 0.55%.

Slant medium: Sodium Glutamate 1%, peptone 0.8%, sodium-chlor 0.5%, sal epsom 0.1%, potassium primary phosphate 0.05%, yeast extract paste 0.5%, Carnis Bovis seu Bubali cream 0.5%, agar 2%.

The activation moral A Kun that goes down to posterity breathes out pseudomonas on aseptic slant medium; Culture temperature 30 ± ℃, the moral A Kun Kazakhstan pseudomonas after the activation is inoculated in the seeding tank that aseptic shake-flask seed liquid culture medium is housed cultivates 30 ± 1 ℃ of jar temperature; Tank pressure 0.05～0.08MPa, ventilation 20～24m ³/ h, when OD has a net increase of 0.5～0.6, endpoint pH is that 8.0～9.0 continuous 6 hours streams when constant add aspartic acid, control pH value, minimum is 6.0, and when pH value is 6.0, continuous 1 hour is constant, and completion is fermented.

1M ³The seeding tank fermented liquid can transform 2.5 tons of aspartic acids, transformation efficiency 95%, L-L-Ala extraction yield 98%.

Claims (1)

1. a method of producing the L-L-Ala comprises the steps:

The activation moral A Kun that goes down to posterity breathes out pseudomonas on aseptic slant medium; Moral A Kun Kazakhstan pseudomonas after the activation is inoculated in the aseptic shake-flask seed liquid culture medium to be cultivated; When OD has a net increase of 0.5～0.6, the pH value be 8.0～9.0 continuous when constant stream add aspartic acid, control pH value is minimum to be 6.0; When the pH value is 6.0, accomplish fermentation when constant continuously;

Said shake-flask seed liquid culture medium is Sodium Glutamate 1%, peptone 0.8%, sodium-chlor 0.5%, sal epsom 0.1%, potassium primary phosphate 0.05%, corn dry powder 0.55%.

Said slant medium is Sodium Glutamate 1%, peptone 0.8%, sodium-chlor 0.5%, sal epsom 0.1%, potassium primary phosphate 0.05%, yeast extract paste 0.5%, Carnis Bovis seu Bubali cream 0.5%, agar 2%.