Professional Documents
Culture Documents
[111
3,867,555
Newell et al.
154]
3,681,195
3,718,541
3,725,075
[75]
Filed:
Louis, Mo.
Nov. 29, 1972
[ 57]
8/1972
2/1973
4/1973
ABSTRACT
[51]
[581
[56]
References Cited
UNITED STATES PATENTS
3,268,412
3,585,179
3,615,654
3,634,194
8/1966
6/1971
10/1971
1/1972
3,867,555
1
IYP.
'
ber.
further advantage is that the non-protein soluble cyto
Cell rupture, extraction of solubles, and processabil
plasmic constituents of the cell can be recovered and
ity
are affected by pH, temperature, time, solids con
processed to a valuable product.
centration, and homogenizer efficiency. Our usual
Our process is comprised of the following steps: pro
duction of yeast cells, rupture of the cells, separation 60 method of measuring the extent of cell rupture is to de
termine the amount of nitrogen that remains soluble,
of the insoluble cell wall fragments from the soluble cy
i.e., the % N Extractability = 100 X
toplasmic fraction, treatment of the soluble fraction
with alkali, recovery'of the low nucleic acid protein by
precipitation and centrifugation, vacuum concentra
tion, and drying.
65
DETAILED DESCRIPTION
Yeast cells (biomass) is produced by methods known
3,867,555
3
through a Manton-Gaulin homogenizer to a chilled re
Saccharomyces cerevisiae
TABLE lA
15
Separation
pH of
Extraction
Solids
Content
7
8
2.5
2.5
3
3
2.5
10
ll
2.5
2.5
3
3
Good
Good
84
Good
82
80
Good
Good
9.5
2.5
70
Good
9.5
9.5
9.5
9
[0
ll
[2
9
10
1l
12
2.5
2.4
2.5
2.4
2.4
2.4
2.4
6.9
6.9
6.9
6.9
2
3
4
3
3
3
3
3
3
3
3
83
89
91
83
82
78
85
64
59
54
41
Good
Good
Good
Good
Medium
Medium
Medium
Medium
Poor
Poor
Poor
Time
C.
No. of
7: Nitrogen
Content
(min. )
Temp.
Passes
Extractability
5.0
6.0
7.0
8.5
9.5
6.0
6.5
7.5
8.5
9.5
3-4
34
3-4
3-4
3-4
3-4
3-4
3-4
3-4
3-4
30
30
30
30
30
60
60
60
60
60
25
25
25
25
25
60
60
6O
6O
6O
3
3
3
3
3
3
3
3
3
3
36
79
93
93
96
42
33
30
73
90
Solids
pH
Sar'clmromyces cererisiae
Solids
Time
C.
No. of
7: Nitrogen
pH
Content
( min. )
Temp.
Passes
Extractability
9.5
9.5
9.5
9.5
9.5
9.5
9.5
9.5
9.5
9.5
9.5
9.5
9.5
4.0
9.l
4.8
3.1
3.1
3 l
3-4
3-4
3-4
3-4
3-4
3-4
3-4
3-4
3-4
30
30
30
30
30
5
20
30
60
5
20
30
60
30
25
25
25
25
25
50
50
50
50
60
60
60
60
25
3
3
3
2
l
3
3
3
3
3
3
3
3
3
83
84
92
80
63
91
93
96
96
93
94
9]
90
33
65
3,867,555
6 .
Incubation Temperature
NaOH [N]
1hr.
60C.
2hr.
3hr.
1hr.
80C.
2hr.
4hr.
1hr.
100C.
2hr.
36
0.025
52
67
80
75
63
78
94
0.05
0.075
6
X
23
X
46'
X
69
83
86
86
81
86
90
90
97
93
100
100
0.10
53
82
88
X i
0.15
79
87
91
'
4hr.
'4 Nucleic Acid Hydrolysis = 100 X Total Am, of Acid Whey of Sample/Total Aw, of Acid Whey of an aliquot
of the alkali extract heated for 30 min.; 100C; in 0.2N KOH.
TABLE H13
92 Precipitation Yield 2
,
60C .
NaOH [N]
0
0.025
0.05
1hr.
2hr.
80
81
8 0C.
3hr.
81
lhr.
2hr.
100C.
4hr.
lhr.
2hr.
X'
4hr.
82
78
74
' 74
74
65 -
79
77
74
73
68
69
62
48
48
0.975
68
65
58.
56
' 52
52
0.10
71
66
64
0.15
67
63
60
' X
' X
X '
n 1'" 71 Precipitation Yield = 100 X kjeldahl N content of Alkali Extract-N Content of Acid Wh'ey/Kjeldahl N
Content of Alkali Extract
"" Theoretical yield: a full RNA-1Y1 is obtained in 8091 precipitation yield and contains 10.7 RNA. 74.6% crude
protein. 11 all of the RNA has been hydrolyzed. and is not precipitable, then the precipitation yield cannot be
greater than 686; .
per 100 ml.). The residue is removed by centrifugation. 45 Temperatures greater and lesser than those given in
The AM of the suitably diluted supernatant is mea-'
Table II have been investigated. At 30C.,incubati0n
for 2 hours at pH 10, 10.5 and 1 1.0 did not present suf
sured. The extinction coefficient of 31.7 A260 ml/mg. is
ficient RNA hydrolysis. Incubation at 120C. for .1 hour
at pHs 4.5, 5.5, 6.5, 7.5, 8.5 and 9.5, followed by pre
50
ture.
DESCRIPTION OF DRAWING
The drawing is a schematic ?ow sheet of the process
of this invention.
3,867,555
EXAMPLE 2
'
EXAMPLE 1
'
is l to 4 hrs.
'
EXAMPLE 3
Nutritional Quality of Unfractionatcd Candida utili.~ and of isolated Yeast Protein (lYP)
Isolated yeast proteins were produced from Candida ulilix in accordance with the process set forth in
Examples 1 and 2. In addition, a full RNA-IYP was prepared according to Example 1 except that the
nucleic acid reduction step was omitted. The isolated yeast proteins and the unfractionated yeast were
Product Composition
Level of
7: dsb
Corrected
Nucleic Acid
Test No.
l
2
Material
Reduction
Step
Corrected
Protein
Nucleic
Acid
Protein in
the Diet
PER
C. utilis
Unfractionated
C. utilis
None
41.6
7.6
10.0
1.47
Homogenate
None
33.4
5.4 -
10.0
1.63
None
42.0
6.7
10.0
1.70
None
None
40.5
62.4
6.0
10.2
10 .0
10.0
1.62
1.60
C . utilis
C . utilis
Homogenate
Full RNA-IYP
Homogenate
3,867,555
9
10
Test No.
Nucleic Acid
Reduction
Step
Material
Product Composition
7: dsb
Correctedz
Nucleic
Protein
Acid
Level of
Corrected
Protein in
the Diet
PER"'
6
7
8
9
10
11
12
Full RNA-IYP
Low RNA-1Y1
Low RNA-IYP
Low RNA-IYP
Low RNA-IYP
Low RNA-IYP
Low RNA-lYP +
None
MSE
LTHA
LTHA
LTHA
LTHA
65.4
67.4
68.0
68.0
68.0
70.0
11.5
1.4
1.6
1.6
1.6
2.0
10.0
10.0
10
12.5
15.0
10
1.55
1.64
0.51
0.71
0.83
0.41
13
14
Meth. (4)
Low RNA-IYP
Casein (ANRC)
LTHA
HTLA
X
70.0
71.2
X
2.0
2.0
X
10
10
7.5
1.87
0.71
2.40
15
Casein
16
17
Casein
Casein
10.0
2.50
X
X
X
X
X
X
12.5
15.0
2.31
2.10
' In accordance with our copending application Newell ct al. A Process 01' Making Yeast Protein Isolate Having Reduced
Nucleic Acid Content". This process uses malt sprout extract (MSE) to hydrolyze the nucleic acid in the alkali extract before
the protein is separated. This produces an IYP having low RNA content.
"-" ('orrccted protein = 6.25 (Total N by Kjcldahl - ,1 Nucleic Acid Content/6.13)
'l" The Protein [Equivalence Ratio (PER) was measured by WARF. Inc. calculated to a PER = 2.5 for ANRC casein at 10%
'
Table IV shows the essential amino-acid composition 40 total lysine. Threonine was not affected as evidenced ,
of the isolated yeast protein products. The full RNA
by the comparison of column and mba values. Cystine,
lYP (Tests No. 5 and No. 6) and the low RNA-lYP
TABLE IV
Essential Amino Acid Composition of Isolated Yeast Protein from Candida utilis I
Amino Acid
Method
of
Detn.
Lysine
FAQ)
Amino Acid)
Pattern
Reqd by the
Growing Rat
Column
8.46
9.56
8.70
9.60
8.12
7.86
8.0
4.2
9.0
Lysine
Lysine
Methionine
C stine
mba
Availab1e"
mba
mba
9.05
7.95
1.35
0.68
X
X
1.32
1.28
X
X
1.36
1.34
9.00
7.92
1.67
0.25
X
X
1.41
. 0.56
X
X
1.30
0.42
X
X .
2.84
0.52
X
X
2.2
2.0
X
X
3.4
T reoninc
Threonine
Tryptophan
Valine
Column
mba
Column
Column
5.25
5.40
1.37
6.41
5.42
X
1.35
6.46
5.40
X
1,53
6.60
5.00
4.84
1.59
6.29
4.95
X
1.47
6.38
5.05
X
1.39
6.34
3.52
X
1.27
7.12
2.8
X
1.4
4.27
5.0
X
1.1
5.5
1.6
3,867,555
11
12
TABLE IV Continued
Essential Amino Acid Composition of Isolated Yeast Protein from Candida urilis
Grams of Amino Acid per 100 Grams Corrected Lrotein
Method of RNA Reduction
Methodm
of
Amino Acid
Phenylalanine
Leucine
None
No.
Detn.
cam};
Column
'
. MSE
No.
No.
533
5.26
55157
9.29
9.48
9.68
LTHA
No.
No.
11
9.41
9.34
Amino Acid)
HTLA
ANRCi
No. .
Casein
13
" 5.32
FAO
' -
'
5.02
Pattern
V. -
Reqd by the
Growing Rat
'28
"
452'
9.26
9.65
4.8
7.0
lsoleucine
Column
575
6.06
6.05
5.88
5.70
5.80
4.85
4.2
5.5 '
Tyrosine
Histidine
Column
Column
6.39
2.51
4.59
3.20
4.85
2.67
4.43
2.80
4.42
2.54
4.74
2.38
5.55
3.20
2.8
X
3.0
2.5
"The amino acid contents were determined by WARF. Inc. by the amino acid analyzer (column), by microbiological assay (mba). and by USP XVlll, 947 (1970)
(Available).
mMalt Sprout Extract - See footnote No. l of Table 111.
debris fraction,
0. Hydrolyzing the nucleic acid with alkali at a pH of
about 9.5 to about 12.5 and a temperature of about
50C. to about 120C. for less than 4 hours,
35
'
>
0% to about 1% ?ber.
for up to 4 hours.
'
>
'
'
5 minutes,
e. Removing the yeast insolubles at a temperature not .