JPH0686686A - Production of epsilon-poly-l-lysine - Google Patents

Abstract

PURPOSE:To provide the method for efficiently producing an epsilon-poly-L-lysine, enabling the semi-continuous production or continuous production of the epsilon-poly-L- lysine without lysing the cells of a bacterium by employing the immobilized cells of the bacterium. CONSTITUTION:Immobilized cells produced by immobilizing the genus Streptomyces organism having an epsilon-poly-L-lysine-producing ability are used under an aerobic condition for the production of the epsilon-poly-L-lysine, and the produced epsilon-poly-L-lysine is recovered. The epsilon-poly-L-lysine-producing bacterium can repeatedly be used, and the bacterial cells are readily removed and recovered, thereby permitting to economically supply the epsilon-poly-L-lysine.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention uses .epsilon.
The present invention relates to a method for producing poly-L-lysine.

[0002]

2. Description of the Related Art ε-Poly-L-lysine is, for example, Streptomyces albulus subsp. Lysinopolym .
erus ) No. It is already known that it can be obtained by culturing 346 (JP-A-53-72896).
issue). The substance is a homopolymer of L-lysine,
It is a polymer compound in which the amino group at the ε-position of lysine is linearly bonded to the carboxyl group of the adjacent L-lysine through a peptide bond. Since the substance is a polymer of L-lysine which is an essential amino acid, it is highly safe and has a high cation content, and thus has unique physical properties. Therefore, by utilizing these properties, applications such as toiletry products, cosmetics, feed additives, agricultural chemicals, food additives, and electronic materials have been developed.

The conventional method for producing ε-poly-L-lysine is as follows:
I went as follows. That is, Streptomyces having the ability to produce ε-poly-L-lysine
The cells of the genus are cultured under aerobic conditions, and after the growth of the cells is observed, the pH is adjusted to around 4 and the culture is continued. Then ε-
After separating the cells of the culture solution containing poly-L-lysine by centrifugation or filtration, for example, a basic anion exchange resin treatment (JP-A-2-20295) or a cation exchange resin treatment (JP-A-2-92927), etc. It was obtained by purification by.

[0004]

[SUMMARY OF THE INVENTION Streptomyces having ε- poly -L- lysine producing ability by conventional methods (Stre
cells of the genus ptomyces ) are cultured under aerobic conditions to obtain ε-poly-
When L-lysine is accumulated in the culture medium, it cannot be reused because the culture medium becomes highly viscous and the cells lyse. Further, when centrifugation or filtration is performed to separate the cells, it takes a long time because the cells are lysed. For example, it takes about 32 hours to remove bacterial cells from a culture solution of 10 m ³ using a filtration device having a filtration area of 10 m ² , which is a major obstacle to production. Further, since the cells are lysed, it is difficult to collect only the culture solution, and it is difficult to perform semi-continuous culture in which a new medium is added. The above factors prevent reduction of the production cost of ε-poly-L-lysine. Therefore, if it is possible to increase the viscosity of the culture solution by the bacterial cells and suppress the lysis of the bacterial cells, the production cost of ε-poly-L-lysine can be reduced. As a result of extensive studies on means for achieving this object, it was found that the above problems can be solved by immobilizing a microorganism having an ability to produce ε-poly-L-lysine, and the present invention was completed. did.

As is clear from the above description, the object of the present invention is to increase the viscosity of a culture solution by using immobilized cells when producing ε-poly-L-lysine using a microorganism.
Immobilized cells isolated by suppressing lysis of the cells, facilitating recovery of the culture solution containing ε-poly-L-lysine and purification of ε-poly-L-lysine from the recovered culture solution Is repeatedly used to semi-continuously or continuously produce ε-poly-
The present invention provides a novel and efficient method for producing ε-poly-L-lysine used as a catalyst for L-lysine production.
In the present invention, semi-continuous production means that a medium containing a substrate or a raw material is added to a reactor containing immobilized cells, and the entire amount of this medium is replaced with a new medium at regular time intervals. Is to repeatedly produce ε-poly-L-lysine.

[0006]

The present invention has the following (1).
(2) It has the configurations of (3), (4) and (5). (1) A method for producing ε-poly-L-lysine, which comprises culturing, under aerobic conditions, an immobilized microbial cell on which a microorganism capable of producing ε-poly-L-lysine is immobilized. (2) ε-poly-using the immobilized bacterial cell described in (1) above
A method for producing ε-poly-L-lysine, characterized in that the method for producing L-lysine is a semi-continuous method or a continuous production method. (3) The method for producing ε-poly-L-lysine according to the above (1) and (2), wherein the microorganism capable of producing ε-poly-L-lysine is a bacterium belonging to the genus Streptomyces. (4) The method for producing ε-poly-L-lysine according to the above (1) and (2), wherein the microorganism having the ability to produce ε-poly-L-lysine is immobilized by an adsorption method, a crosslinking method or an encapsulation method. . (5) The ε-poly-L according to the above (1) and (2), wherein the microorganism having the ability to produce ε-poly-L-lysine is immobilized by a combination of an adsorption method and a crosslinking method or an adsorption method and an inclusion method. -Method for producing lysine.

The structure and effects of the present invention will be described in detail below. As the microorganism used in the present invention, any microorganism can be used as long as it has the ability to produce ε-poly-L-lysine. For example, Streptomyces albulus subspecies
us subsp. lysinopolymerus ) No. 346 (JP-A-53
-72896) and [epsilon] -poly-L-lysine in significant production mutant No. 11011A-1 (Japanese Patent Laid-Open No. 63-4907)
5), Streptomyces
Noursei ) (JP-A-1-187090) and the like are known.

The ε-poly-L-lysine-producing bacterium used in the present invention can be immobilized by a general method and application of a general method. For example, adsorption method, crosslinking method,
Immobilized cells can be prepared by using the entrapment method, the covalent bond method, etc. as a single immobilization method, or the immobilization method in which two kinds of these methods are combined can be applied. Of these, it is easy to immobilize and stable activity is obtained,
Furthermore, the adsorption method and entrapment method, which can reactivate when the activity decreases, can be preferably used. In the encapsulation method, it is common to enclose the bacterial cells in a polymer gel such as polyacrylamide gel, polyurethane gel, and photocrosslinkable resin. However, in the present invention, since the product ε-poly-L-lysine is a basic substance, it is preferable not to use an acidic polymer gel that adsorbs this substance, such as carrageenan gel or alginic acid gel. . As the adsorption method, a method of immobilizing it on a porous material or a non-woven fabric is particularly preferable. As the porous material, for example, porous ceramics, porous glass, cellulose foam, sintered metal porous body or polyvinyl chloride, polyethylene, polypropylene, polystyrene,
It is also possible to use one prepared from polyurethane or the like.

For encapsulation by the above polyacrylamide gel, for example, acrylamide monomer, N, N'-methylenebisacrylamide as a cross-linking agent and viable cells are suspended in a buffer solution, and ammonium persulfate as a polymerization initiator is suspended in the buffer solution. Then, β-dimethylaminopropionitrile as a polymerization accelerator is added, and a polymerization reaction is carried out at room temperature for about 30 minutes to obtain immobilized cells.

Further, the thermoplastic resin is generally 150 to
By heating at a temperature of 250 ° C. for 0.5 to 5 hours, a sintered body is obtained, and if a suitable mold is used, sintered porous bodies having various shapes can be obtained. In addition, the desired porosity can be obtained by changing the conditions such as the porosity, the size of the pores, the strength, etc., such as the sintering temperature, the sintering time, the resin filling rate, the amount of the thickener or the amount of water added. As the above-mentioned other porous substances, all commercially available products and processed products which can adsorb the present producing bacteria can be used. The porous substance can be immersed in a buffer solution containing viable cells to adsorb the microorganisms, or the microorganisms and the porous substance can be contacted from the start of the culture and adsorbed as the culture progresses. Is obtained.

The immobilized microbial cell having the ability to produce ε-poly-L-lysine obtained by the above-mentioned method contains glucose and glucose in a usual medium for producing ε-poly-L-lysine or a buffer solution whose pH is appropriately adjusted. A culture solution containing ε-poly-L-lysine can be obtained by culturing under aerobic conditions in a medium containing ammonium sulfate or L-lysine. The immobilized cells are removed from the main culture solution, and ε-poly-L-lysine is purified using an ion exchange resin. In addition, as the pH during culture, the used ε-
Microorganisms capable of producing poly-L-lysine are ε-poly-
It is sufficient that L-lysine can be produced, preferably pH 4 to 7.

[0012]

According to the method of the present invention, microbial cells are not lysed during culturing, immobilized cells can be easily removed, and ε-poly-L-lysine solution containing no bacterial cells can be obtained in a short time. Can be collected in
It enables efficient production of ε-poly-L-lysine. Further, since the immobilized cells and the culture solution can be easily separated, after the production of ε-poly-L-lysine, the culture solution is recovered and a new medium is added to the immobilized cells to repeatedly produce ε. -Poly-
Semi-continuous ε-poly-, such as producing L-lysine
By producing L-lysine, or continuously discharging the culture solution and adding a new medium, continuous production of ε-poly-L-lysine can be performed. From the above, ε-poly-L-lysine is efficiently industrially produced at low cost for a long time,
It is extremely useful in that it can be supplied to fields such as food additives, agricultural chemicals, and pharmaceuticals.

[0013]

EXAMPLES The present invention will be described below based on examples.
The present invention is not limited to this. Example 1 For immobilization, glucose 50 g / L, ammonium sulfate 10 g / L, yeast extract 5 g / L, potassium dihydrogen phosphate 1.36 g / L, disodium monohydrogen phosphate 1.58
g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.
A medium (pH 6.8) 50 ml consisting of 04 g / L and ferrous sulfate 0.03 g / L was placed in a 500 ml Sakaguchi flask, sterilized by a conventional method, and Streptomyces albulus was inoculated, 36 at 30 ° C
The culture was shaken for a period of time to obtain a preculture liquid. After completion of the culture, cells were collected by centrifugation and washed twice with sterilized water (wet cell weight 4.2 g).

The immobilized cells were prepared as follows. Acrylamide 1.68 g, N, N′-methylenebisacrylamide 0.09 g, L-lysine hydrochloride 0.6
7 g and the bacterial cells obtained by the above method were suspended in 4.8 ml of Tris-hydrochloric acid buffer (pH 7.2), and a 5% β-dimethylaminopropionitrile aqueous solution 1.
1 ml and 1.1 ml of 1% potassium persulfate aqueous solution were added,
After leaving it to stand at room temperature for 30 minutes under N ₂ saturation condition to cause gelation,
This gel was molded into a 5 mm square, and then the gel was thoroughly washed with Tris-hydrochloric acid buffer solution to obtain 11.2 g of polyacrylamide gel-immobilized cells.

The polyacrylamide gel-immobilized cells were added to 40 ml of a medium (pH 4) containing glucose 50 g / L, L-lysine 10 g / L, and citric acid 20 g / L, 30
The culture was carried out with shaking at ℃. 2 days later, ε-poly-L in the culture solution
-The amount of lysine was determined to be 0.23 g / L.

Example 2 75 g of polyacrylamide gel-immobilized cells prepared by the same method as in Example 1 was used as a medium for culturing ordinary .epsilon.-poly-L-lysine-producing bacteria, glucose 50 g / L, ammonium sulfate 10 g / L. , Yeast extract 5 g / L, potassium dihydrogen phosphate 1.36 g / L, disodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L, ferrous sulfate 0.03 g / L (pH
4.2) 50 ml was added and shake culture was performed at 30 ° C. The amount of ε-poly-L-lysine in the culture medium after 2 days was
It was 0.20 g / L.

Example 3 100 g of polyacrylamide-immobilized cells prepared by the same method as in Example 1 above was used as a medium (pH 4) of glucose 50 g / L, L-lysine 10 g / L and citric acid 20 g / L.
The mixture was added to 1 L, and aeration stirring culture was carried out at 30 ° C. while adjusting the pH to 4 using a 1.5 L mini jar fermenter. Ε-Poly-L in the culture medium after 48 hours of culture
-The amount of lysine was 1.5 g / L. Then, glucose and L-lysine were sequentially added to continue the culture. After 125 hours, the amount of ε-poly-L-lysine was 10 g / L.
The culture solution was centrifuged (3000 G, 20 minutes) to remove bacterial cells, and the permeability of the centrifuged supernatant was measured at 660 nm. The permeability was 0.008, and the cells were completely removed.

Comparative Example 1 Glucose 50 g / L, ammonium sulfate 10 g / L,
Yeast extract 5g / L, potassium dihydrogen phosphate 1.36g
/ L, disodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L,
Medium consisting of ferrous sulfate 0.03 g / L (pH 6.8)
50 ml is put in a 500 ml Sakaguchi flask, sterilized by a conventional method, and then Streptomyces albus ( Streptom)
yces albulus ) was inoculated and shake-cultured at 30 ° C. for 36 hours to prepare a preculture liquid. This preculture solution was inoculated into a 1.5-liter mini jar fermenter containing 1 liter of the same medium as described above, and aeration-agitation culture was performed at 30 ° C. After culturing, after the pH of the culture solution was lowered to about 4, the pH was adjusted to 4.2 using an aqueous sodium hydroxide solution, and the glucose concentration was sequentially added to 50 g / L, and the culture was continued. The amount of ε-poly-L-lysine in the culture medium after culturing for 125 hours was 12 g / L. The culture solution was centrifuged (3000 G, 20 minutes) to remove bacterial cells, and the permeability of the centrifuged supernatant was measured at 660 nm. The transparency is
At 0.282, the removal of bacterial cells was insufficient. From the above results, it was confirmed that bacterial cells can be easily removed by using the immobilized bacterial cells of the present invention.

Example 4 The bacterial cells (wet bacterial cell weight: 2.0 g) that were cultured and collected in the same manner as in Example 1 were suspended in 2 ml of 0.1 M phosphate buffer solution and shown in [Chemical formula 1]. 3 g of a urethane prepolymer having a structure (a molecular weight of about 3000) was added and mixed under stirring to form a gel, which was then molded into a 5 mm square to obtain 6.7 g of a polyurethane gel-immobilized bacterium. The polyurethane gel-immobilized cells were added to 40 ml of a medium (pH 4) containing glucose 50 g / L, L-lysine 10 g / L and citric acid 20 g / L at 30 ° C.
The cells were shaken and cultured. Ε in the culture medium after 2 days and 3 days
-The amount of poly-L-lysine was quantified. The amounts of ε-poly-L-lysine were 0.31 g / L and 0.47 g / L, respectively.

[0020]

[Chemical 1]

Example 5 After culturing for 3 days under the same conditions as in Example 4, centrifugation (30
The supernatant was removed with 20 g of 00 G), and glucose 50 g / L, L-lysine 10 g / L, and citric acid 20 were newly added.
40 ml of a g / L medium (pH 4) was added and shake culture was performed at 30 ° C. 2 days after adding new medium,
The amount of ε-poly-L-lysine in the culture solution was 0.75 g / L.
Met. From this, it was confirmed that the immobilized bacterial cell of the present invention can be used for semi-continuous culture.

Example 6 A photo-crosslinkable resin ENT-2000 (manufactured by Kansai Paint Co., Ltd.) was prepared by suspending the cells (wet weight 2.0 g) cultured and collected in the same manner as in Example 1 in 5 ml of sterilized water and sterilized by an autoclave. 40% of a 15% aqueous solution of 0.1) and a polymerization initiator of 0.
And 008 g were added and mixed. Then, the suspension was irradiated with ultraviolet rays of 365 nm for 30 minutes while stirring to prepare a cell-immobilized gel. This cell-immobilized gel is 5m
It was molded into m-square to obtain 53 g of immobilized microbial cells.

The photo-crosslinked resin-immobilized bacterial cells were treated with glucose 5
0 g / L, L-lysine 10 g / L, citric acid 20 g / L
Was added to 100 ml of the medium (pH 4) and cultured at 30 ° C. with shaking. The amount of ε-poly-L-lysine in the culture solution was quantified after 2 days and 3 days. The amounts of ε-poly-L-lysine were 0.44 g / L and 0.57 g / L, respectively.

Example 7 Cells precultured in the same manner as in Example 1 were collected by centrifugation and washed with sterilized water (wet cell weight 4.0 g). Glucose 50g / L, ammonium sulfate 10g
/ L, yeast extract 5 g / L, potassium dihydrogen phosphate 1.
36 g / L, disodium monohydrogen phosphate 1.58 g /
L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04
medium consisting of g / L and ferrous sulfate 0.03 g / L (pH
6.8) 2 g of a polyurethane sintered porous body having a diameter of 5 mm was placed in 30 ml, and gently shake-cultured at 30 ° C. After 30 hours, the bacterial cells immobilized with the polyurethane sintered porous body were collected by centrifugation. 50 g / L glucose, 10 g / L L-lysine, and 2 citric acid
Suspend in 50 ml of medium (pH 4) containing 0 g / L, and
Shaking culture was performed at 0 ° C. Ε in the culture medium after 2 days of culture
-The amount of poly-L-lysine was 0.62 g / L.
In addition, this culture solution is centrifuged (3000 G, 20 minutes)
Then, the transmittance of the supernatant was measured at 660 nm. The transparency is
At 0.009, the cells were completely removed.

Example 8 Glucose 50 was added to a bubble aeration type reactor having a capacity of 400 ml.
g / L, ammonium sulfate 10 g / L, yeast extract 5 g
/ L, potassium dihydrogen phosphate 1.36 g / L, disodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L, ferrous sulfate 0.03 g / 100 ml of medium (pH 6.8) consisting of L
And 30 g of block-shaped ceramic (manufactured by NGK Insulators) are put in and sterilized by a standard method, and then Streptomyces albulus is taken from the slant culture medium for preserving the bacteria 1
Platinum loops were inoculated, and the air flow rate at 30 ° C was 0.5 L / min. At 50
Aeration culture was performed for an hour. The amount of ε-poly-L-lysine in the culture medium after 50 hours was 0.3 g / L. Then, only the culture solution was collected, and glucose 50 g / L, L-lysine hydrochloride 10 g / L, citric acid 20 g / L, potassium dihydrogen phosphate 1.36 g was newly added to the immobilized cells in the reactor. / L, sodium monohydrogen phosphate 1.58
g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.
100 ml of a medium containing 04 g / L and ferrous sulfate 0.03 g / L (pH 4) was added, and aeration culture was carried out at 30 ° C. for 48 hours. The amount of ε-poly-L-lysine in the culture medium after 48 hours was 3.7 g / L. Further, collecting the culture solution,
The operation of newly adding a medium and performing aeration culture at 30 ° C. for 48 hours was repeated 3 times. As a result, the production amount of ε-poly-L-lysine in each repeated cycle was 4.5 g /
L, the second time was 4.0 g / L, and the third time was 5.2 g / L. From this, it was confirmed that ε-poly-L-lysine can be semi-continuously produced by repeating the medium exchange using the immobilized bacterium of the present invention.

Example 9 Glucose 50 was added to a bubble-aeration type reactor having a capacity of 400 ml.
g / L, ammonium sulfate 10 g / L, yeast extract 5 g
/ L, potassium dihydrogen phosphate 1.36 g / L, disodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L, ferrous sulfate 0.03 g / 100 ml of medium (pH 6.8) consisting of L
And 1 g of Cellulose Foam Micro Cube FN-S03 (manufactured by Sakai Engineering Co., Ltd.) were sterilized by a conventional method, and then Streptomyces ablus ( Streptomyces a
lbulus ) is inoculated with 1 platinum loop from a slant culture medium for bacterial storage, and 30
Aeration rate of 0.5 L / min. Aeration culture was carried out for 50 hours. The amount of ε-poly-L-lysine in the culture solution injection after 50 hours was 0.33 g / L. Then, only the culture solution was collected, and glucose 50 g / L, L-lysine hydrochloride 10 g / L, citric acid 20 g / L, potassium dihydrogen phosphate 1.36 g was newly added to the immobilized cells in the reactor. / L,
Sodium monohydrogen phosphate (1.58 g / L), magnesium sulfate (0.5 g / L), zinc sulfate (0.04 g / L), ferrous sulfate (0.03 g / L, pH 4) (100 ml) was added to the medium at 48 ° C at 30 ° C. Aeration was performed for an hour. The amount of ε-poly-L-lysine in the culture medium after 48 hours was 2.9 g / L. Furthermore, the culture solution is recovered, and a new medium is added to
The operation of performing aeration culture at 0 ° C. for 48 hours was repeated 3 times. As a result, the amount of production of ε-poly-L-lysine at each repetition was 3.0 g / L at the first time and 2.5 g / L at the second time.
L was 2.0 g / L for the third time. From this, it was confirmed that semi-continuous production of ε-poly-L-lysine is possible by repeating the medium exchange using the immobilized bacterium of the present invention.

Comparative Example 2 Bacteria were cultured in Example 8 without adding a ceramic carrier. That is, glucose of 50 g / L, ammonium sulfate of 10 g / L, yeast extract of 5 g / L, potassium dihydrogen phosphate of 1.36 g / L, and disodium monohydrogen phosphate of 1.5 were added to a 400-mL volume bubble-reacting reactor.
Put 100 ml of medium (pH 6.8) consisting of 8 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L, and ferrous sulfate 0.03 g / L, and sterilize by a conventional method, and then streptomyces.・ Albrus ( Streptomyces a
lbulus ) is inoculated with 1 platinum loop from a slant culture medium for bacterial storage, and 30
Aeration rate of 0.5 L / min. Aeration culture was carried out for 50 hours. The amount of ε-poly-L-lysine in the culture medium after 50 hours was 0.25 g / L. Then, the culture solution is centrifuged (6000 G, 15 minutes) to collect bacterial cells, and 50 g / L of glucose, 10 g / L of L-lysine hydrochloride, 20 g / L of citric acid, and phosphorus are newly collected from the collected bacterial cells. Potassium dihydrogen acid 1.36 g / L, sodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L, ferrous sulfate 0.03 g / L (pH
100 ml of the medium of 4) was added, and the mixture was aerated at 30 ° C. for 48 hours. The amount of ε-poly-L-lysine in the culture medium after 48 hours was 1.3 g / L. Further, the culture solution was centrifuged (6000 G, 15 minutes) to collect the bacterial cells, the above-mentioned new medium was added again, and aeration culture was carried out at 30 ° C. for 48 hours. The amount of ε-poly-L-lysine in the culture solution injection after 48 hours was 0.7 g / L. From this result, the ε-poly-
In the semi-continuous production of L-lysine, it was confirmed that the productivity of ε-poly-L-lysine decreases as the number of times is repeated, and the semi-continuous production is difficult.

Example 10 Glucose 50 was added to a bubble aeration type reactor having a capacity of 400 ml.
g / L, ammonium sulfate 10 g / L, yeast extract 5 g
/ L, potassium dihydrogen phosphate 1.36 g / L, disodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L, zinc sulfate 0.04 g / L, ferrous sulfate 0.03 g / 100 ml of medium (pH 6.8) consisting of L
And 30g of block-shaped ceramic (manufactured by NGK Insulators) are added and sterilized by a conventional method, and then Streptomyces albulus ( Streptomyces albulus ) is added from a slant culture medium for bacterial preservation 1
Platinum loops were inoculated, and the air flow rate at 30 ° C was 0.5 L / min. At 50
Aeration culture was performed for an hour. The amount of ε-poly-L-lysine in the culture medium after 50 hours was 0.3 g / L. Then, glucose 50 g / L, L- was newly added into the reactor.
Lysine hydrochloride 10 g / L, citric acid 20 g / L, potassium dihydrogen phosphate 1.36 g / L, sodium monohydrogen phosphate 1.58 g / L, magnesium sulfate 0.5 g / L,
Zinc sulfate 0.04 g / L, ferrous sulfate 0.03 g / L
Add 2 ml of (pH 4) medium and remove the culture solution.
Continuous production of ε-poly-L-lysine was carried out at a rate of hr for 300 hours. The total amount of ε-poly-L-lysine produced in 300 hours was 2.2 g. From this, it was confirmed that continuous production of ε-poly-L-lysine can be easily performed by continuously exchanging the medium using the immobilized bacterium of the present invention.

Claims (5)

[Claims] 1. A method for producing ε-poly-L-lysine, which comprises culturing an immobilized microbial cell on which a microorganism capable of producing ε-poly-L-lysine is immobilized under aerobic conditions. 2. An ε-using the immobilized bacterial cell according to claim 1.
A method for producing ε-poly-L-lysine, wherein the method for producing poly-L-lysine is a semi-continuous method or a continuous production method. 3. The microorganism capable of producing ε-poly-L-lysine is a bacterium belonging to the genus Streptomyces.
And the method for producing ε-poly-L-lysine according to claim 2. 4. The method for producing ε-poly-L-lysine according to claim 1 or 2, wherein immobilization of a microorganism having an ability to produce ε-poly-L-lysine is carried out by an adsorption method, a crosslinking method or an inclusion method. . 5. The ε-poly-L according to claim 1 or 2, wherein immobilization of a microorganism having the ability to produce ε-poly-L-lysine is carried out by a combination of an adsorption method and a crosslinking method or an adsorption method and an inclusion method. -Method for producing lysine.