JP2793720B2 - Method for producing monoacetyl polyamine - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a process for producing monoacetyl polyamine, which comprises hydrolyzing diacetyl polyamine in the presence of diacetyl polyamine amide hydrolase. More specifically, the present invention relates to a method for producing monoacetylpolyamine, which comprises reacting diacetylpolyamine with microbial cells having a hydrolytic ability of diacetylpolyamine, a crude enzyme extract obtained from the cells, or a purified enzyme. .

[0002]

2. Description of the Related Art Diacetylpolyamine amide hydrolase is an enzyme that hydrolyzes diacetylpolyamine to produce monoacetylpolyamine, and is, for example, an enzyme that catalyzes the following reaction.

Diacetyl putrescine + H ₂ O → monoacetyl putrescine + CH ₃ COOH

Heretofore, there has been little knowledge about amidohydrolases acting on diacetylpolyamine, and thus no industrially applicable technology has been established.

In addition, when monoacetyl polyamine is synthesized from diacetyl polyamine by organic chemistry to obtain pure monoacetyl polyamine, a large amount of free polyamine is by-produced, and the operation of separation and purification is complicated. And the yield is low.

[0006]

In view of the above background, it has been desired to find an enzyme which acts on diacetylpolyamine to produce monoacetylpolyamine, diacetylpolyamine amide hydrolase.

[0007]

Means for Solving the Problems The inventors of the present invention have made intensive studies to solve the above-mentioned drawbacks and found that diacetylpolyamine amide hydrolase, an enzyme acting on diacetylpolyamine to produce monoacetylpolyamine, was introduced into microorganisms. It has been found that monoacetyl polyamine can be produced by reacting diacetyl polyamine amide hydrolase with diacetyl polyamine, thereby completing the present invention.

[0008] That is, the present invention is a method for producing monoacetyl polyamine, which comprises hydrolyzing diacetyl polyamine in the presence of diacetyl polyamine amide hydrolase.

As the diacetyl polyamine used in the present invention, known diacetyl polyamines can be used without any particular limitation. Specific examples of particularly preferred ones are diacetyl-1,2.
-Diaminoethane, diacetyl-1,3-diaminopropane, diacetylputrescine, diacetylcadaverine, diacetyl-1,6-diaminohexane, diacetyl-1,8-diaminooctane, diacetylspermidine, diacetylspermine and the like.

The microorganisms having a diacetylpolyamine amide hydrolase-producing ability suitably used in the present invention are as follows.

The genus Alcaligenes, the genus Pseudomonas, the genus Esherichia
a) genus, Aerobacter, Serratia (S
erra-tia), Streptomyces (Storeptomyces),
Genus Nocardia, Mycobacterium (Myco
bacterium), Aspergillus, Mucor, Fusarium, Chaetomium, Debaryomyce
s) Genus, genus Candida, Rhodotorula (Rhodo
microorganisms belonging to the genus torula) or the genus Trichosporon, among which genus Alcaligenes and Pseudomonas
s) genus, Fusarium genus, Candida (Ca
A microorganism belonging to the genus ndida) or the genus Trichosporon is preferably used.

[0012] As strains of these microorganisms, for example, Escherichia coli (Esherichia coli IFO 12713), Aerobacter cloacae IAM 122
1), Serratia marcescens
IFO 3054), Alcaligene
s faecalis IFO 14479), Alcaligenes faecalis IFO 13111), Alcaligenes faecalis
Species (Alcalige-nes SP. IFO 14130), Pseudomonasa aureofaciens (Pseudomonasaureofaci)
eus IFO 3521), Pseudomonas sinzanta (Pseud)
omonas Synzantha IFO 3913), Pseudomonas Synzantha IFO 3912, Pseudomonas aeruginosa IFO 3
080), Pseudomonas
Chlororaphis IFO 3904), Streptomyces grisues IFO 12875), Streptomyces coelicolor IFO 128
54), Streptomyces fragee (Storeptomyces fr)
adiae IFO 12773), Streptomyces Alps (Store
ptomyces albus IFO 13014), Nocardiaerythropolis IFO 12682, Mycobacterium phlei (Mycobacterium phleiIFO 13160),
Mycobacterium smegmatis
egmatisIFO 3154), Aspergillus niger
lus niger IFO6661), Aspergillus oryzae (Asper)
gillus oryzae IFO 30105), Mucor javanicus IFO 4570, Ketomium globosum (Chaetomium globosum ATCC 6205), Fusarium oxysporum IFO 7190, Fusarium solani (Fusarium solani Seomyces) hansenii IFO 0794),
Candidatropicalis IFO
0007), Candida guillierm
ondiiIFO 0454), Rhodotorula r
ubra IFO 0870), Rhodotorula r
ubra IFO 1101), Trichosporon Kutanium (Tri-c)
hosporon cutaneum IFO 1198) and Trichosporon cutaneum IFO 0173. Of these strains, Alcaligenes faecalis IFO 14479, Pseudomonas chlororaphis IFO 390
4), Pseudo-monas synxan
tha IFO 3913), Fusarium oxysporum
um oxyspo-rum IFO 7190), Fusarium Solani (Fusa
rium solani IFO 9955), Candida guilliermondii IFO 0454, Trichosporon cutaneum IFO 1198, Trichosporon cutaneum (Trichosporon cutaneum
IFO 0173) and the like are preferably used.

As a medium used for culturing each of the above microorganisms, a known medium is used. For example, glucose, honey, carbon source such as soluble starch, meat extract, yeast extract, nitrogen source such as polypeptone, and potassium phosphate monobasic, potassium phosphate dibasic, sodium chloride,
It is not particularly limited as long as it contains inorganic salts such as magnesium sulfate.In addition to these components, diacetyl polyamines such as diacetyl putrescine and diacetyl cadaverine, monoacetyl putriscine, and monoacetyl-
It is important to include a monoacetyl polyamine such as 1,6-diaminohexane.

The diacetylpolyamine or monoacetylpolyamine may be added to the medium from the beginning or may be added during the culturing. The medium may be in the form of a liquid or a solid. The culture method may be any of stationary culture, shake tow culture, and aeration and agitation culture. For large-scale culture, liquid culture using aeration and agitation is suitable. The culture temperature is 15 to 40
° C. Preferably, it is performed at 20 to 35 ° C.

In order to carry out the production method of the present invention, the culture solution obtained as described above, the cells collected from the culture solution or the processed product of the cells, for example, washed cells, dried cells, bacteria, etc. Acts on body mass, autolysed cells, sonicated cells, cell extracts, or purified enzymes on diacetylpolyamine in inorganic or organic solvents that do not inhibit the reaction, preferably in aqueous solvents By doing so, a monoacetyl polyamine can be produced. Alternatively, a cell, a crude enzyme extract, or a purified enzyme immobilized by a commonly used method for immobilizing cells or enzyme can also be used.

The concentration of diacetylpolyamine added is about 10
There is no particular limitation as long as it is at least mM and does not inhibit the reaction. The temperature at which the enzyme acts may be any temperature at which the enzyme does not die, and is usually about 10 to about 50 ° C, preferably about 25 to about 40
° C. The pH is from 6 to 10, preferably from 7 to 9.

When diacetylpolyamine is added to the culture, it is preferably added during the logarithmic growth phase. The concentration to be added may be about 10 mM or more as long as it does not inhibit the reaction.

When the cells are used, the cells in the logarithmic growth phase and stationary phase cultured in the above medium are collected by centrifugation, washed with physiological saline, etc., and then washed with 50 mM phosphate buffer (pH 7.0).
0) etc., and diacetylpolyamine is added to about 10
It may be added at a concentration that does not inhibit the reaction at mM or more.

The monoacetylpolyamine can be recovered from the thus obtained culture solution or reaction solution by a known method such as ion exchange column chromatography and crystallization.

Among the above methods, the cells obtained by culturing in the presence of diacetylpolyamine are separated, the cells are crushed by ultrasonic waves or the like, and the resulting crude enzyme extract is subjected to ammonium sulfate fractionation, dialysis, and column chromatography. It is preferable to use an enzyme preparation purified by chromatography, gel filtration, or the like. Among such specimens, Alcaligenes faeca-
lis IFO 14479), and Candida giramondi (Ca
ndida guilliermondii IFO 0454), and a diacetylpolyamine amide hydrolase produced according to Example 18 or 20 described later is specifically shown below as an enzyme preparation.

Alcaligenes faeca-li was added to a medium containing 0.2% diacetylputrescine.
s IFO 14479) or Candida Giramondi (Candid
a guilliermondii IFO 0454)
After culturing at 28 ° C. for 24 to 48 hours in a fermenter, the cells were separated from the culture solution, and the crude enzyme extract obtained by crushing the cells after ultrasonic crushing or dynomill was used as an ion exchange column The physicochemical properties of diacetylpolyamine amide hydrolase purified by chromatography, gel filtration, hydrophobic column chromatography and the like are as follows.

[Diacetylpolyamine amide hydrolase derived from the genus Alcaligenes] 1. Action The amide bond of diacetylpolyamine is hydrolyzed to produce monoacetylpolyamine. 2. Substrate specificity The relative activity of this enzyme for each substrate at a concentration of 10 mM is shown in Table 1 when the activity for cyacetylptrescine is expressed as 100. 3. Optimum pH: pH 8.5 to 9.5 (Fig. 1). 4. pH stability When stored at 30 ° C for 30 minutes, pH stability is 9 at pH 7.0-9.0.
It has a residual activity of 0% or more (FIG. 2).

Table 1 {Relative activity of substrate (%)} ─────────────────────────────── diacetyl-1,2-diaminoethane 5.7 diacetyl-1,3-diaminopropane 67.7 diacetylputrescine 100 diacetylcadaverine 78.7 diacetyl-1,6-diaminohexane 96.0 diacetyl-1,8-diaminooctane 21.0 diacetylspermidine 5.5 diacetylspermine 3.8───── ────────────────────────────── * N ¹ -acetylsperlysine formation 5. Optimum temperature at pH 9.0 for 15 minutes It is 37-42 ° C in the reaction (Fig. 3). 6. Temperature stability Under treatment conditions of pH 7.5 and 30 minutes, it has a residual activity of 90% or more at a temperature up to 53 ° C., and is completely inactivated by a treatment at 67 ° C. for 30 minutes (FIG. 4). ).

[Candida sp.-derived diacetylpolyamine amide hydrolase] 1. Action The amide bond of diacetylpolyamine is hydrolyzed to produce monoacetylpolyamine. 2. Substrate specificity The relative activity of this enzyme for each substrate at a concentration of 10 mM is shown in Table 2 when the activity for cyacetylptrescine is expressed as 100.

Table 2 {Relative activity of substrate (%)} ───────────────────────────────Diacetyl-1,2-diaminoethane 91.9 Diacetyl-1,3-diaminopropane 17.8 diacetylputrescine 100 diacetylcadaverine 65.0 diacetyl-1,6-diaminohexane 39.4 diacetyl-1,8-diaminooctane 26.3 diacetylspermidine 10.0 diacetylspermine 15.1 ────────────────────────────── * N ¹ -acetylspermidine formation 3. Optimum pH: pH 7.0 to 8. 0 (FIG. 5). 4. pH stability When stored at 30 ° C. for 30 minutes, it has a residual activity of 90% or more at pH 5.5 to 11.3 (FIG. 6). 5. Optimum temperature: 37-42 ° C in a reaction at pH 7.5 for 15 minutes (Fig. 7). 6. Temperature stability Under the treatment conditions of pH 7.5 and 30 minutes, it has a residual activity of 90% or more at temperatures up to 32 ° C., and is completely inactivated by treatment at 55 ° C. for 30 minutes (FIG. 8). ).

The method for measuring the enzyme activity of diacetylpolyamine amide hydrolase and the method for displaying the enzyme activity in the present invention are as follows.

1.0 ml of 20 mM diacetylputrescine
And acylpolyamine amide hydrolase (JP-B-59)
-7435) 6.8 units, putrescine oxidase
0.28 units, peroxidase 0.9 units, 2.4
-0.1 M Tris-HCl buffer (pH 8.0) containing 0.75 mg of dichlorophenol and 0.06 mg of 4-aminoantipyrine 1.
The reaction is carried out by mixing 50 μl of a test sample containing 0 ml of diacetylpolyamine amide hydrolase, reacting the mixture at 30 ° C. for 1 hour, and measuring the absorbance at 510 nm.

The enzyme activity value is expressed as the amount of the enzyme which produces 1 μmole of monoacetyl putrescine per minute as 1 unit (μmole / min).

[0029]

Examples 1 to 16 0.5% polypeptone, 0.01% yeast extract, 0.1% salt, 0.2% diacetylputrescine, 0.05% potassium monophosphate, 0.15% phosphorus 10 ml of a medium of potassium diacid (pH 7.0) was dispensed into a large test tube and autoclaved at 120 ° C. for 15 minutes. One loopful of various microorganisms shown in Table 2 was inoculated from the slant culture, Shaking culture was performed at 28 ° C for 24-120 hours. After completion of the culture, the cells were collected by centrifugation and washed with 5 ml of 10 mM phosphate buffer (pH 7.5).
The suspension was suspended in 5 ml of 10 mM phosphate buffer (pH 7.5) and sonicated (180 W, 5 to 10 minutes) under ice-cooling. The sonicated solution was centrifuged to obtain a crude enzyme extract. To 0.2 ml of the crude enzyme extract was added 1.0 ml of a 100 mM phosphate buffer (pH 7.5) and 0.8 ml of 20 mM diacetylputrescine to give 30 minutes.
The reaction was carried out at a temperature of 3 ° C. for 3 hours. After the reaction was stopped by adding 0.2 ml of 50% trichloroacetic acid, the produced monoacetylputrescine was quantified by high performance liquid chromatography. The results are shown in Table 2. The produced monoacetyl putrescine (μmole) in the table was obtained at a reaction temperature of 30 ° C. for 3 hours.
The amount of monoacetyl putrescine produced by the action of 2 ml of the crude enzyme extract, and the activity per 1 ml of the culture medium is the enzyme activity of the crude enzyme extract converted to 1 ml of the original culture solution.

[0030]

[Table 1]

Example 17 0.5% polypeptone, 0.01% yeast extract, 0.1% salt, 0.05% potassium monophosphate, 0.15% dipotassium phosphate, 0.2% diacetyl 0.5 L of a medium (pH 7.0) containing putrescine and 0.02% defoamer was placed in a 2 L Erlenmeyer flask, autoclaved at 120 ° C. for 20 minutes, and then Alcaligenes faecalis IFO 144 was added.
79) was inoculated. After shaking culture at 28 ° C. for 48 hours, this culture solution was previously added to a medium 2 having the same composition as described above.
0 L was added to a sterilized jar fermenter, and main culture was performed. Culture conditions: 28 ° C, stirring speed: 100r
After culturing for 48 hours at 20 L / min with aeration at 20 pm, the culture was centrifuged to collect the cells.

About 200 g (wet cell weight) of the obtained cells
Was suspended in 1 L of 10 mM phosphate buffer (pH 7.5), and the suspension was disrupted with an ultrasonic disrupter (220).
W, 80 min). The crushed liquid was centrifuged using a centrifuge to obtain a crude enzyme extract, and the activity was measured by the above-mentioned enzyme activity measuring method. As a result, the enzyme activity of the crude enzyme extract was 0.71 U / ml. From the enzyme activity value, it was found that monoacetylputrescine 2.1 μmole was produced in 0.05 ml of the crude enzyme extract at 30 ° C. for 1 hour.

Example 18 In Example 17, Alcaligenes faecalis (A
lcaligenes faecalis IFO 14479) was previously purified from a crude enzyme extract obtained from a 20 mM phosphate buffer (pH
The DEAE-cellulose column (60
0 ml) and eluted with a linear concentration gradient of sodium chloride (0.1 to 0.7 M). After collecting the active fractions and desalting by ultrafiltration, ammonium sulfate was added to 20%,
Then, butyl Toyopearl 650 previously equilibrated with a 20 mM phosphate buffer (pH 7.5) containing 20% ammonium sulfate was used.
M (manufactured by Tosoh Corporation) column (100 ml), and a reverse linear concentration gradient elution of ammonium sulfate (20 to 0%) was performed. The active fractions were collected, desalted by ultrafiltration, and then subjected to gel filtration on a Sephacryl S-300 (Pharmacia) column (1.8 L). The active fraction was collected and then E was pre-equilibrated with 20 mM phosphate buffer (pH 7.5).
AH-Sepharose (Pharmacia) column (40
ml) and eluted with a linear concentration gradient of sodium chloride (0 to 0.6M). The active fractions were collected, desalted by ultrafiltration, and then adsorbed on a column (70 ml) of DEAE-Toyopearl 650S (manufactured by Tosoh Corporation) previously equilibrated with 20 mM phosphate buffer (pH 7.5). (0 to 0.35 M). The active fractions were collected, desalted by ultrafiltration, and ammonium sulfate was added to a concentration of 15%.
Phenyl-equilibrated with 0 mM phosphate buffer (pH 7.5)
Sepharose 4B (Pharmacia) column (100
ml), and a reverse linear concentration gradient elution of ammonium sulfate (15-0%) was performed. The active fractions were collected and desalted by ultrafiltration to obtain 37 ml of a purified enzyme solution containing 38.5 mg of protein. The physicochemical properties of the enzyme are as described above. Except that 0.05 ml of this purified enzyme solution was used, the hydrolysis reaction of diacetylputrescine was carried out in exactly the same manner as in Example 17. As a result, the enzyme activity of the purified enzyme solution was 13.0 units / ml, and the specific activity was 12.5 units / mg-.
It was a protein. Further, from the enzyme activity value, it was found that monoacetyl putrescine 39 μmole was produced in 0.05 ml of the purified enzyme solution at 30 ° C. for 1 hour.

Example 19 Candida guilliermondiiIF
O 0454), a crude enzyme extract was obtained and the enzyme activity was measured in exactly the same manner as in Example 17, except that the culture time was 24 hours, and the cells were disrupted by Dynomill. Was. As a result, the enzyme activity of the crude enzyme extract was 0.11 units / ml. In addition, 3
One hour at 0 ° C., 0.05 ml of the crude enzyme extract produced 0.32 μmole of monoacetyl putrescine.

Example 20 In Example 19, Candida giramondi (Candida) was used.
ida guilliermondii (IFO 0454) was preliminarily purified from a crude enzyme extract obtained from the cells of 20 mM phosphate buffer (pH 7.5).
Was adsorbed on a DEAE-cellulose column (1.5 L) equilibrated in step, and a linear concentration gradient elution of sodium chloride (0 to 0.5 M) was performed. The active fractions were collected and desalted by ultrafiltration, and ammonium sulfate was added to a concentration of 25%.
20 mM phosphate buffer containing 5% ammonium sulfate (pH
The mixture was adsorbed to a phenyl-Sepharose 4B (Pharmacia) column (100 ml) equilibrated in 7.5), and a reverse linear concentration gradient elution of ammonium sulfate (25 to 0%) was performed. The active fractions were collected, desalted by ultrafiltration, and then subjected to gel filtration on a Sephacryl S-300 (Pharmacia) column (1.8 L). The active fraction was collected and then E was pre-equilibrated with 20 mM phosphate buffer (pH 7.5).
AH-Sepharose (Pharmacia) column (40
ml), and a linear concentration gradient elution of sodium chloride (0 to 0.25 M) was performed. Active fractions were collected, desalted by ultrafiltration, and then DEAE-Toyopearl 650S (manufactured by Tosoh Corporation) previously equilibrated with a 20 mM phosphate buffer (pH 7.5).
The mixture was adsorbed on a column (10 ml), and a linear gradient elution of sodium chloride (0 to 0.2 M) was performed. The active fractions were collected, desalted by ultrafiltration, and then 10 mM phosphate buffer (pH 7.
A DEAE-cellulose column (20) equilibrated in 5)
ml) and eluted with a linear concentration gradient of sodium chloride (0 to 0.2 M). The active fractions were collected and desalted by ultrafiltration to obtain 7.0 ml of a purified enzyme solution containing 1.9 mg of protein. The physicochemical properties of the enzyme are as described above. Except that 0.05 ml of this purified enzyme solution was used, a hydrolysis reaction of diacetylputrescine was performed in exactly the same manner as in Example 17. As a result, the enzyme activity of the purified enzyme solution was 3.6 units / ml, and the specific activity was 13.2 units / mg-protein. Further, according to the enzyme activity value, it was found that 11 μmole of monoacetyl putrescine was produced at 30 ° C. for 1 hour with a purified enzyme activity of 0.05 ml.

Example 21 0.05 ml of the purified enzyme solution obtained in Example 18 was mixed with 0.1 ml of a 100 mM solution of various diacetylpolyamines and 0.85 ml of 0.1 M carbonate buffer (pH 9.0). Add 3
The reaction was performed at 7 ° C. for 15 minutes. 50% trichloroacetic acid
After adding 1 ml to stop the reaction, the produced monoacetylpolyamine was quantified by high performance liquid chromatography. The results are shown in Table 3.

Table 3 Diacetylpolyamine Monoacetylpolyamine production (10 ³ × μmole) ────────────────────────────────── diacetyl-1,2-diaminoethane 11.7 diacetyl- 1,3-diaminopropane 135 diacetylputrescine 902 diacetylcadaverine 710 diacetyl-1,6-diaminohexane 866 diacetyl-1,8-diaminooctane 189 diacetylspermidine 49.6 ^* diacetylspermine 34.3 ──────────────────────────── * N ¹ -acetylspermidine formation

Example 22 0.05 ml of the purified enzyme solution obtained in Example 20 was combined with 0.1 ml of a 100 mM solution of various diacetylpolyamines and 0.85 ml of a 0.1 M phosphate buffer (pH 7.5). Add 3
The reaction was performed at 7 ° C. for 15 minutes. 50% trichloroacetic acid
After adding 1 ml to stop the reaction, the produced monoacetylpolyamine was quantified by high performance liquid chromatography. The results are shown in Table 4.

Table 4 {Diacetylpolyamine Monoacetylpolyamine production (10 ³ × μmole) ────────────────────────────────── diacetyl-1,2-diaminoethane 682 diacetyl-1, 3-diaminopropane 132 diacetylputrescine 742 diacetylcadaverine 482 diacetyl-1,6-diaminohexane 292 diacetyl-1,8-diaminooctane 195 diacetylspermidine 47.2 ^* diacetylspermine 112 ──────────────────────── * N ¹ -acetylspermidine formation

Example 23 0.5 g of each of the cells obtained in Examples 17 and 19 was added to 20%
The solution was added to 2.0 ml of 50 mM phosphate buffer (pH 7.5) containing diacetylputrescine, reacted at 30 ° C. for 20 hours, and centrifuged to remove cells. 50 in the supernatant
After 0.2 ml of trichloroacetic acid was added to terminate the reaction, the produced monoacetylputrescine was quantified by high performance liquid chromatography. The results are shown in Table 5.

Table 5 ────────────────────────────────── Microorganism name Monoacetyl putrescine ( μmole) ────────────────────────────────── Alcaligenes faecalis 680 Candida giramondi 536 ───── ─────────────────────────────

Examples 24 to 39 Diacetylputrescine was hydrolyzed in exactly the same manner as in Examples 1 to 16, except that the culture solutions of the various microorganisms obtained in Examples 1 to 16 were used. The results are shown in Table 6.
The amount of monoacetyl putrescine formed in the table was 30 ° C., 3
This is the amount produced by the action of 0.2 ml of the culture solution in the reaction over time.

[0043]

[Table 2]

[0044]

Industrial Applicability According to the present invention, monoacetylpolyamine useful as an intermediate for medical and agricultural chemicals can be produced under mild conditions and in good yield. In addition, the present invention provides a technique for industrially utilizing an amide hydrolase that acts on diacetylpolyamine.

The present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

[Brief description of the drawings]

FIG. 1 is a view showing a pH activity curve of diacel polyamine amide hydrolase derived from the genus Alcaligenes.

FIG. 2 shows the pH stability of the enzyme.

FIG. 3 is a view showing a temperature activity curve of the enzyme.

FIG. 4 shows the temperature stability of the enzyme.

FIG. 5 is a view showing a pH activity curve of diacetylpolyamine amide hydrolase derived from Candida.

FIG. 6 shows the pH stability of the enzyme.

FIG. 7 is a diagram showing a temperature activity curve of the enzyme.

FIG. 8 shows the temperature stability of the enzyme.

────────────────────────────────────────────────── ─── Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI (C12P 13/00 C12R 1: 185) (C12P 13/00 C12R 1:01) (C12P 13/00 C12R 1: 465) (C12P 13/00 C12R 1: 365) (C12P 13/00 C12R 1:32) (C12P 13/00 C12R 1:69) (C12P 13/00 C12R 1: 785) (C12P 13/00 C12R 1:77) (C12P 13/00 C12R 1:72) (C12P 13/00 C12R 1: 645) (C12P 13/00 C12R 1:19) (C12P 13/00 C12R 1:43) (56) References JP-A-2-101056 ( JP, A) JP-A-59-25692 (JP, A) JP-A-56-151494 (JP, A) APPL. MICROBIOL. , VOL. 10 (1962), p. 401-406 (58) Field surveyed (Int. Cl. ⁶ , DB name) C12P 13/00-13/24 CA (STN)

Claims (2)

(57) [Claims] 1. A method for producing monoacetyl polyamine, comprising hydrolyzing diacetyl polyamine in the presence of diacetyl polyamine amide hydrolase. 2. A diacetylpolyamine according to the genus Alcaligenes, Pseudomonas, Escherichia, Aerobacterium, Serratia, Streptomyces, Nocardia, Mycobacterium, Aspergillus, Mucor, Fusarium, Ketomium, Monoacetylpolyamine characterized in that it hydrolyzes in the presence of a culture solution, microorganisms, or microbial cells of a microorganism belonging to the genus selected from the group consisting of Debaryomyces, Candida, Rhodotorula, and Trichosporone Manufacturing method.