Patented'Mer-3,1949
- 2,463,784
/
UNITED STATES, PATENT OFFICE
sm'rnon or raeraama PENTONIC scms
raonr rau'rosas
Lewis B. Lockwood, Peoria, 111., assignor to the
United States of America as represented by the
Secretary of Agriculture
No Drawing. Application August 9, 1946,
'
Serial No. 689,350
14 Claims. (01. 195-41)
(Granted under the act 0! March 3, 1883, as
1
amended April 30, 1928; 370 O. G. 757)
2
This application is made under the act or
stirrers may be used as suitable vessels in which
March 3, 1883, as mended by the act of April
to carry out the process.
30, 1928, and the invention herein described, it
Considerable latitude is possible in the selec
patented, may be manufactured and used by or
tion of the nutrients to be used in the conduct
for the Government of the United States of 5 of this invention.‘ Thus, the KHaPO4, MgSO4,
America for governmental purposes without the
‘and urea may be varied in quantity, or may be
payment to me of any royalty thereon.
omitted, and corn steep liquor, vegetable decoc
This invention relates to a method of prepartions, liver extract, or any of the wide variety of
ing pentonic acids from the corresponding aldo‘nutrient materials now marketed for the culture
pentoses by fermentation with bacteria of the 10 of micro-organisms, may be substituted for the
genus Pseudomonas.
There are eight possible
- dehydrated yeast extract.
aldo-pentoses. They are d-arabinose, l-arabi-
The following examples illustrate representa
nose, d-lyxose, l-lyxose, d-ribose, l-rigose, Ig-xyl-
tive procedures used in practicing this invention.
ose, and l-xylose. Of these, only -ara nose,
l-arabinose, d-ribose, and d-xylose are of com- “5
mon occurrence.
This invention relates, more
EXAMPLE I
The non-saccharine constituents of all of the
particularly, to the oxidation of these four pen-
culture solutions were the same. They constitut
toses to the corresponding pentonic acids, respectively, d-arabonic acid, l-arabonic acid, d-ribonic
ed urea, 02 8111-; KHiPQi. 0-6 gm: MgS04HHz0,
9-25 8111-; dehydrated yeast extract, 0.5 gm.; soy
acid, and d-xylonic acid.
'
20 Pea-n 011. 3 drops P91‘ 100 ml. culture; and CaCOz
Fermentative oxidation of- sugars has a num"1 excess of the quantity necessary to neutralize
ber 'of advantages over chemical oxidation of
the maximum quantity of pentonic acid which
‘ sugars in that costly reagenm are not required’
could theoretically be formed from the pentose. ,
better yields of oxidized products may be obtained
The culture solutions were aerated by bubbling
under suitable conditions, and the microbial oxi- 25 air through them at the rate of 100 ml. air per 100
dation processes are more speci?c, resulting in
In]- mash Pe1'_ minute- The temperature was
fewer side reactions leading to cgntaminating
maintained at 30° C'. d-Arabinose was added to
products which interfere with recovery_
the basal culture solution, and 100 ml. portions
The Oxidation of arabinose to arabonjc acid
Of were inoculated with bacteria Of the species:
was reported by Bertrand (Compt. Rend. Acad. 3" Pseudomonas fmgii, ‘P. graveolen, P- synrantha.
Sci" Paris, 1279129431 (1393), who used acetic
and P. vendrelli. Analyses of mashes, made aft
acid bacteria Bertrand (compt; Rend_ Acml
er seven days, gave the data contained in Table I.
Sci., Paris, 127:124-127 (1898)), also reported
TABLEI
the oxidation of xylose to xylonic acid by acetic
.
_
v
‘
acidbacteria. The use of bacteria of the genus 35 4'8 grams of d'ambmose in 100 ml- culture 801%“
Pseudomonas for pentose oxidation has not been
“on
previously reported in the literature. The use
of Pseudomonas instead of acetic acid bacteria
Grams
Weightyleld
o?ers the advantages that the process is of wider
applicability, since acetic acid bacteria do not 40
q-mbonic d?f?c
wtgr‘irdtdt?‘fd Miami-100
oxidize some of the pentoses which are readily
culture
oxidized by bacteria of the Pseudomonas genus,
'
and since pure synthetic nutrients may be used,
Psmdommmgfmgii
(1.5313059
'
____________ __-_
20
2M
‘although
trients may
thebeaddition
made inofsome
cmdecases
nitrogenous
where these
nu- 45'Pseudomonaa
lli?ggm wag
marezuzajj _______ _jj
2-53
173
1512
will not interfere with recovery. It is well known
that acetic acid bacteria require the addition of
‘ expensive vitamins if purely synthetic mashes are
to be used_
such additions are not required by
' bacteria
In thisofinvention
the genus
thepseudomonas.
mashes may be aerated,
either at atmospheric pressure or at super-atmospheric pressure by any means known to those
skilled in the art. Rotating drums, or deep ves-
EXAMPLE n
-
l-Arabinose instead of d-arabinose was added
It0 the
100basal
ml- Culture
P01130118Solution
were inoculated
described inwith bac
time Of the Species PSeudOmOMs ?uofeseens,
P- fmyii, 1’- mildenbefyii, P- pu?dd, P- Syn-‘mutila
and P. vendrelli. Analyses made after six days
sels equipped with disperser stones, agitators, or 05 gave the data presented in Table II.
2,463,784
3
2. A process of making d-arabonic acid coni
prising inoculating a nutrient medium containing
d-arabinose with bacteria of the genus
Pseudomonas, and aerating the medium, a
TABLE II
4.0 grams of l-arabinose per 100 ml. culture solu
tion
.
Grams,
Weight yield
l-arabonic
kgxgg?c
acid produced acid “100
per 100 ml.
grgms
culture
l-arabinose
Paeudomonas ?uorescent 6 ___________ ..
Pseudomonas fragii 25 _____ ._
.
1. 38
28. 2
l. 75
35. 7
1. 41
28. 8
Pseudomonas mildenbcrgii 21..
Pseudomonas putida 13.. _._
2. 34
47. 8
Pacudomonas sunzantha 79..
Pacudomoncs vendrelli 23 _____________ ..
. 86
2. 22
17.5
45. 4
neutralizing agent for the pentonic acid formed
being present.
3. A process of making d-ribonic acid compris
ing inoculating a medium containing d-ribose
with bacteria of the genus Pseudomonas, and
10 aerating the medium, a neutralizing agent for the
pentonlc acid formed being present.
.4. A process of making d-xylonic acid com
prising inoculating a medium containing d-xylose
with bacteria of the genus Pseudomonas, and
15 aerating the medium, a neutralizing agent for the
pentonic acid formed being present.
EXAMPLE III
cl-Ribose, instead of d-arabinose, was added to
the basal culture solution described in Example I.
100 ml. portions were inoculated with bacteria of
of the species Pseudomonas ?uorescens, P.
5. The process of claim 1 in which the bacteria
is Pseudomonas ?uorescens.
6. The process of claim 1 in which the bacteria
0 is Pseudomonas mildenbergii.
'7. The process of claim 2 in which the bacteria
graveolens, P. mildenbergii, P. ovalis, P.-putrija
ciens, P. synzcantha, and P. uendrelli. Analyses
8. The process of claim 3 in which the bacteria
is Pseudomonas ?uorescens.
9. The process of claim 4 in which the bacteria
is Pseudomonas jragii.
made after nine days gave the data in Table III.
TABLE III
5.9 grams d-ribose per 100 ml. culture solution
Pseudomonas fragii 25 ________________ ..
Pseudomoriaa mildenberaii 21.- ..
Paeudomimas ovalis 8 ______ ..
Pseudo'mzmas putri/acicnc 76
Paeudomonac aynzantha 79.
Pseudo-menus cendrelli 23...
-
is Pseudomonas ?uorescens.
10. The process of claim 4 in which the bacteria
is Pseudomonas mildenbergii.
11. The process of claim 1 in which the
Grams d- Weight yield
neutralizing agent is a calcium compound.
ribonic acid
grams d
produced
ribonic acid
12. The process of claim 1 in which the
per 100 ml. per 100 grams
neutralizing agent is calcium carbonate and the
culture
d-ribose
temperature is maintained at about 30° C., the
calcium carbonate being in excess of the quantity
1.08
18. 4
0. 58
9. 9 35 necessary to neutralize the maximum theoretical
0. 41
7.0
yield of pentonic acid from the pentose.
0.81
14.8
13. A process of converting d-xylose to d
l. 69
28.8
..
2. 89
49. 3
Pscudomonas ?uorescent 6. ...'....... ..
4.11
70.0
EXAIVIPLE IV
d-Xylose, instead of d-arabinose, was added to
the basal culture solution described in Example I.
100 ml. portions were inoculated with bacteria
of the species Pseudomonas J‘iuorescens, P. fragii,
P. graveolens, P. mildenbergz'i, P. ovalis, and P.
putida. Analyses made after three days gave the
data presented in Table IV.
.
‘TABLE IV
6.7 grams d-mylose per 100 ml. culture solution
Grams
d-xylonic
acid roducc
er
Weight yield
grams
xylonic acid in high yields, comprising inoculat
ing a nutrient medium containing d-xylose with
40 Pseudomonas
KH2PO4, MgSOl, dehydrated yeast extract, and
calcium carbonate in excess of the quantity
necessary to neutralize the maximum theoretical
yield of d-xylonic acid from d-xylose.
14. A process of making a pentonic acid com
prising inoculating a nutrient medium contain
ing an aldo-pentose with bacteria of the genus
Pseudomonas, and aerating the medium, the said
50 medium containing urea, KHzPO4, MgSO4, de
hydrated yeast extract, and calcium carbonate
in excess of the quantity necessary to neutralize
> 1the maximum theoretical yield of pentonicacid
d-xylonic
from the pentose.
acid per 100
100 m .
grams
culture
d-xylose
?uorescens, and aerating the
medium, the said medium containing urea,
-
LEWIS B. LOCKWOOD.
REFERENCES CITED
Pacudomonas ?uorescent 6 ........... ..
Pscudomcmas I wait 25 ............. -.
4. 59
1. 89
68. 5
28. 2
Pacudo'mmm graveolem 14...
Paeudo'monas mildmberaii 2i
3. 78
4. 87
55. 5
72.9
Paeudomimcs ovalia 8. ....
Puudomonus putida 13. ..
2. 66
4. 40
39. 7
65. 7
The following references are of record in the
.
60 file of this patent:
UNITED STATES PATENTS
Name
Date
Number
2,356,581
Having. thus described my invention, I claim:
1. A process of making a pentonic acid com
prising inoculating a nutrient medium containing‘
an aldo-pentose with bacteria of the genus
Pseudomonas, and aerating the medium, a
neutralizing agent for the pentonic acid formed
being present.
65
Grandel __________ __ Aug. 22, 1944
[OTHER REFERENCES
Lockwood et al., J. Bact., July 1941, page 54.
Chemical Abstracts, vol. 34, 7321, '7 and 8
. (1940), citing V.‘ V. Pervozvankii-Microbiology
(U. S. S. R.) 8, N0. 2, 149-159 (1939) , and N0. 3-4,
70 339-52.