Download ENZYME TECHNOLOGY

Enzyme Technology, January 1981
PROGRAM
SRI INTERNATIONAL
Menlo Park, California
Abstract
Process Economics Program Report No. 139
ENZYME TECHNOLOGY
(January 1981)
The report presents the technologies of enzyme production, recovery, purification, immobilization, and application. The process
economics for these technologies are illustrated by:
l
The production of a bran mold and a bran-amylase product by a
surface cultivation process*
l
The production of a bacterial alpha-amylase preparation by a
submerged cultivation process.
l
The immobilization of an enzyme by an entrapment/cross-linking
process.
s
The application of an immobilized enzyme in the production of
high fructose syrup.
.
Included also is a brief review of some of the patents and publications
on enzyme-catalyzed chemical conversions.
PEP '79 WSF
Enzyme Technology, January 1981
Report No. 139
ENZYME TECHNOLOGY
by WING SIEN FONG
0
0a
0m
I
January 1981
I
A private report by the
PROCESS
ECONOMICS
Menlo Park, California
PROGRAM
94025
Enzyme Technology, January 1981
For detailed
referred
marketing
to one of the SRI programs
most major chemicals
States
In addition,
uct,
specializing
is
in marketing
products
produced
in the
and their
derivatives
on a worldwide
basis.
the SRI DIRECTORY
OF CHEMICALPRODUCERS
services
detailed
and plant
and chemical
and the WORLD
PETROCHEMICALS
Program covers
major hydrocarbons
provide
the reader
The CHEMICALECONOMICS
HANDBOOK
Program covers
research.
United
data and information,
lists
for
of chemical
the United
States
ii
producers
by company, prod-
and Western Europe.
Enzyme Technology, January 1981
CONTENTS
1
INTRODUCTION. . . . . . . . . . . . . . . . . . . . . . . . .
1
2
SUMMARY...........................
3
3
INDUSTRY STATUS .......................
11
Product Forms. .......................
Applications. ........................
U.S. Governmental Regulations on the Use
of Certain Enzymes. .....................
Major Producers .......................
12
13
4
TYPICAL PROPERTIES OF ENZYMES . . . . . . . . . . . . . . . .
27
5
ENZYME CATALYSIS
Mechanism .........................
Specificity ........................
Inhibition. ........................
Cofactors .........................
Allosterism and Allosteric Enzymes. ............
Kinetics .........................
Effect of Temperature ...................
Effect of pH ........................
Effect of Immobilization. .................
a
6
0
0
0
......................
16
17
39
39
39
40
41
42
43
49
50
50
COMMERCIAL ENZYME PRODUCTION PROCESSES. . . . . . . . . . . .
59
Animal Enzymes. . . . . .
Pancreatin. . . . . . .
Pepsin . , . . . . . .
Rennin (or rennet), . .
Catalase. . . . . . . .
Plant Enzymes . . . . . .
Malt Amylases . . . . .
Papain . . . . . . . .
Bromelin. . . . . . . .
Ficin . . . . . . . . .
Microorganism Enzymes . .
General Aspects . . . ,
Commercial Processes. .
Surface Cultivation .
Submerged Cultivation
59
60
60
61
61
66
66
67
68
69
74
74
82
85
88
..................
..................
..................
..................
..................
..................
..................
..................
..................
..................
..................
..................
..................
..................
..................
iii
Enzyme Technology, January 1981
CONTENTS
7
ENZYME RECOVERY AND PURIFICATION. ...............
91
91
........................
Precipitation
96
Salting Out .........................
97
Addition of Organic Solvents. ................
98
Addition of High Molecular Weight Substances. ........
98
Addition of Metallic Ions or Complexes. ...........
99
Addition of Specific Agents .................
99
Addition of Carriers. ....................
99
Crystallization ........................
100
Extraction ..........................
100
Electrophoresis ........................
101
Chromatography. ........................
101
Adsorption ..........................
Ion Exchange............................
101
102
Ultrafiltration (Gel Filtration). ...............
103
Biochemical Interaction (Affinity Chromatography) .......
103
Lyophilization (Freeze Drying) ................
..lO 4
Drying ..........................
104
Dialysis. ...........................
104
Activated Carbon Treatment. ..................
104
Concentration .........................
Desalting..................................................
105
Disruption of Microorganism Cells
105
(For Intracellular Enzyme Recovery) ..............
8
ENZYME PRODUCTION BY SURFACE CULTIVATION, A MOLD BRAN
PRODUCT AND A BRAN-AMYLASE PRODUCT ..............
Process Description ......................
MoldBran..........................lO8
Bran-Amylase Product. ....................
Process Discussion. ......................
Cost Estimates, ........................
MoldBran........................115
Bran-Amylase Product. ....................
9.
107
107
112
114
115
116
ENZYME PRODUCTION BY SUBMERGED CULTIVATION,
A BACTERIAL ALPHA-ADYLASE PRODUCT. ..............
129
Process Description ......................
Process Discussion. ......................
Cost Estimates. ........................
129
135
137
IV
Enzyme Technology, January 1981
0
CONTENTS
0
10
a
a
11
12
a
IMMOBILIZATION OF ENZYMES. ..................
147
Immobilization Methods ....................
Adsorption .........................
Entrapment .........................
Microencapsulation .....................
Covalent Binding and Intermolecular Cross-Linking. .....
Immobilization Supports. ...................
Commercial Immobilization Processes. .............
Commercial Applications of Immobilized Enzymes ........
Production of High Fructose Syrup. .............
Production of L-Amino Acids. ................
Production of 6-AminopenicillanicAcid ...........
Hydrolysis of Lactose in Whey. ...............
Commercial Applications of Immobilized Microorganism Cells . .
Enzyme Immobilization Based on Entrapment/
Cross-Linking Technology ...................
Process Description. ....................
Process Discussion .....................
Cost Estimates .......................
148
148
161
164
164
170
172
175
175
175
178
178
181
182
182
187
188
A COMMERCIAL APPLICATION OF AN IMMOBILIZED
ENZYME, HIGH FRUCTOSE SYRUP PRODUCTION ............
195
Review of Processes. .....................
Process Description. .....................
Process Discussion ......................
Cost Estimates ........................
196
204
210
210
CHEMICALS BY ENZYMATIC CONVERSION. ..............
217
Acrylic Acid or Acrylate from a Propionate or
Propionic Acid (457294, 468001). ...............
Acrylsmide or Methacrylamide from Acrylonitrile
or Methacrylonitrile (457318). ................
Desulfurization of Petroleum (457345) ............
Diepoxides from Diolefins (457295) ..............
Epoxides by Olefin Fermentation (457288) ...........
Epoxides and Glycols from Alkenes (457292) ..........
1-Epoxy-Alkanes from 2-20 Carbon u-Olefins or 4-20 Carbon
a,w-Diolefins (457381) ....................
Hydrogen Sulfide from Gypsum (457296). ............
Ketones and Secondary Alcohols from 3-6 Carbon Alkanes and
Ketones from 3-6 Carbon Secondary Alcohols (488006). .....
Others ............................
V
217
218
218
218
219
221
222
224
224
225
Enzyme Technology, January 1981
CONTENTS
APPENDIX A
DESIGN AND COST BASES. . . . . . . . . . . . . . .
APPENDIX B
SOME OF THE MORE COMMON TERMS, ABBREVIATIONS,
227
AND SYMBOLSUSED IN THE ENZYME INDUSTRY.. . . . . 231
APPENDIX C
DEFINITION OF ENZYMATIC ACTIVITY AND SOME
OF THE MORE COMMONLYUSED UNITS. . . . . . . . . . 235
APPENDIX D
CITED REFERENCES
ENZYME REPOSITORIES . . . . . . . . . . . . . , .
239
. . . . . . . . . . . . . . . . . . . . . . . . .
241
PATENT REFERENCES BY COMPANY.
. . . . . . . . . . . . . . . . . . .
Vi
253
Enzyme Technology, January 1981
ILLUSTRATIONS
5.1
l
I
0
Graphical Determination of s and VMAE in the
Michaelis-Menten Equation, . . . . . . . . . . . . . . . . .
47
6.1
Production of Pancreatin . . . . . . . . . . . . . . . . . .
62
6.2
Production of Pepsin . . . . . . . . . . . . . . . . . . . . 63
6.3
Production of Rennin . . . . . . . . . . . . . . . . . . . .
6.4
Production of Catalase . . . . . . . . . . . . . . . . . . . 65
6.5
Productionof Malt Amylases. . . . . . . . . . . . . . . . .
70
6.6
Production of Papain . . . . . . . . . . . . . . . . . . . .
71
6.7
Production of Bromelin . . . . . . . . . . . . . . . . . . .
72
6.8
Production of Ficin
73
6.9
Production of Fungal Enzymes by Surface Cultivation. . . . . 86
6.10
Production of Enzymes by Submerged Cultivation . . . . . . .
8.1
Enzyme Production by Surface Cultivation,
A Mold Bran Product and a Bran-Amylase Product . . . . . . . 257
8.2
Enzyme Production by Surface Cultivation,
A Mold Bran Product
Effect of Bran Price and Changes in Labor, Capital,
or Utilities Cost on Production Cost . . . . . . . . . . . . 119
8.3
Enzyme Production by Surface Cultivation,
A Mold Bran Product
Effect of Product Yield on Production Cost . . . . . . . . . 120
8.4
Enzyme Production by Surface Cultivation,
A Mold Bran Product
Effect of Operating Level and Plant Capacity
on Production Cost . . . . . . . . . . . . . . . . . . . . . 121
8.5
Enzyme Production by Surface Cultivation,
A Bran-Amylase Product
Effect of Bran Price and Changes in Labor, Capital,
or Utilities Cost on Production Cost . . . , . . . . . . . . 125
8.6
Enzyme Production by Surface Cultivation,
A Bran-Amylase Product
Effect of the Amount of Enzyme Concentrate Recovered
from the Dry Bran Weight Loss on Production Cost . . . . . . 126
8.7
Enzyme Production by Surface Cultivation,
A Bran-Amylase Product
Effect of Operating Level and Plant Capacity
on Production Cost . . . . . . . . . . . . . . . . . . . . . 127
. . . . . . . . . . . . . . . . . . . .
vii
64
89
Enzyme Technology, January 1981
ILLUSTRATIONS
.
9.1
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product. . . . . . . . . . . . . .
259
9.2
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Effect of Raw Material Prices on Production Cost . . . . . . 143
9.3
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Effect of Gypsum Dilution on Production Cost . . . . . . . .
144
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Effect of Changes in Utilities, Capital, or Labor Cost
and in Enzyme Yield on Production Cost . . . . . , . . . . .
145
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Effect of Operating Level and Plant Capacity
on Production Cost . . . . . . . . . . . . . . . . . . . . .
146
Some of the More Common Covalent Binding
Reactions Involved in the Immobilization . . . . . . . . . .
166
Flow Diagram for Continuous Production of L-Amino Acid
By Immobilized Aminoacylase. . . . . . . . . . . . . . . . .
177
A Continuous Lactose Hydrolysis Process
Using Immobilized Lactase. . . . . . . . . . . . . . . . . .
180
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology. . . . . . . . . . . . .
261
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology
Effect of Gelatin and Glutaraldehyde Prices and Changes
in Labor, Capital, and Utilities Cost on Conversion Costs. .
192
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology
Effect of Operating Level and Plant Capacity
on Conversion Cost . . . . . . . . . . . . . . . . . . . . .
193
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology
Effect of Soluble Enzyme Cost on Conversion Cost . . . . . .
194
9.4
9.5
10.1
10.2
10.3
10.4
10.5
10.6
10.7
Viii
0
l
Enzyme Technology, January 1981
ILLUSTRATIONS
-
11.1
A Commercial Application of an Immobilized Enzyme,
High Fructose Syrup Production. . . . . . . . . . . . . . .
263
A Commercial Application of An Immobilized Enzyme,
High Fructose Syrup Production
Effect of Changes in Immobilized Enzyme,
Miscellaneous Chemicals, Utilities, Capital,
or Labor Cost on Conversion Cost. . . . . . . . . . . . . .
215
A Commercial Application of an Immobilized Enzyme,
High Fructose Syrup Production
Effect of Operating Level and Plant Capacity
on Conversion Cost. . . . . . . . , . . . . . . . . . . . .
216
12.1
Epoxides of Olefin Fermentation . . . . . . . . . . . . . .
220
12.2
Epoxidation of Unsaturated Hydrocarbons
by Immobilized Cells. . . . . . . . . . . . . . . , . . . .
223
11.2
011.3
iX
Enzyme Technology, January 1981
TABLES
0
2.1
Summary of Economics. . . . . . . . . . . . . . . . . . . .
6
3.1
Applications of Enzymes . . . . . . . . . . . . . . . . . .
14
3.2
Production of Some of the Principal Enzymes in Japan
. . .
19
3.3
Some of the Principal Enzymes Produced
and their Major Producers in Japan. . . . . . . . , . . . .
20
Some of the Principal Enzymes Produced
and their Major Producers in the United States. . . . . . .
22
Some of the Principal Enzymes Produced
and their Major Producers in Western Europe . . . . . . . .
24
4.1
Description of Selected Industrial Enzymes
. . . . . . . .
32
5.1
Properties of Free (Native) and Various
Immobilized Aminoacylases . . . . . . . . . . . . . . . . .
51
Estimated Half-Life for Selected
.Immobilized Enzyme Systems. . . . . . . . . . . . . . . . .
56
Activity and Activity Yield for
Various Immobilized Aminoacylases . . . . . . . . . . . . .
57
Examples of Some of the Product Inducers
and Analog Inducers . . . . . . . . . . . . . . . . . . . .
75
6.2
Examples of Catabolite Repression of Enzymes. . . . . . . .
76
6.3
Some of the Better Known Microorganisms
Used in Enzyme Production . . . . . . . . . . . . . . . . .
79
Patent Summary of Some Microorganism
Enzyme Production Methods . . . . . . . . . . . . . . . . .
83
Patent Summary on Some Enzyme Recovery
and Purification Methods. . . . . . . . . . . . . . . . . .
92
Enzyme Production by Surface Cultivation,
A Mold Bran Product
Major Equipment and Utilities Summary . . . . . . . . . . .
109
Enzyme Production by Surface Cultivation,
A Bran-Amylase Product
Major Equipment and Utilities Summary . . . . . . . . . . .
110
Enzyme Production by Surface Cultivation,
A Mold Bran Product
Total Capital Investment . . . . . . . . . . . . . . . . .
117
Enzyme Production by Surface Cultivation,
A Mold Bran Product
Production Costs. . . . . . . . . . . . . . . . . . . . . .
118
3.4
3.5
5.2
5.3
6.1
6.4
7.1
8.1
8.2
8.3
8.4
xi
Enzyme Technology, January 1981
TABLES
8.5
8.6
9.1
9.2
Enzyme Production by Surface Cultivation,
A Bran-Amylase Product
Total Capital Investment. . . . . . . . . . . . . . . . . .
122
Enzyme Production by Surface Cultivation,
A Bran-Amylase Product
Production Costs . . . . . . . . . . . . . . . . . . . . .
123
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Stream Flows. . . . . . . . . . . . . . . . . . . . . . . .
132
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Major Equipment and Utilities Summary . . . . . . . . . . .
133
9.3
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Total Capital Investment. . . . . . . . . . , . . . . . . . 140
9.4
Enzyme Production by Submerged Cultivation,
A Bacterial Alpha-Amylase Product
Production Costs . . . . . . . . . . . . . . . . . . . . .
141
Patent Summary on Some of the Immobilization Methods
for Enzymes and Microorganism Cells . . . . . . . . . . . .
149
Physical Properties and Applications for
Controlled-Pore Ceramics and Glass. . . . . . . . . . . . .
173
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology
Major Equipment and Utilities Summary . . . . . . . , . . .
185
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology
Total Capital Investment . . . . . . . . . . . . . . . . .
190
Enzyme Immobilization Based on an
Entrapment/Cross-LinkingTechnology
Conversion Costs. . . . . . . . . . . . . . . . . . . . . .
191
11.1
Patent Summary on Some Glucose Isomerization Processes. . .
197
11.2
A Commercial Application of an Immobilized Enzyme,
High Fructose Syrup Production
Major Equipment and Utilities Summary . . . . . . . . . . .
207
10.1
10.2
10.3
10.4
10.5
Xii
r
Enzyme Technology, January 1981
TABLES
11.3
A Commercial Application of an Immobilized Enzyme,
High Fructose Syrup Production
Bases and Assumptions. . . . . . . . . . . . . . . . . . . . 209
11.4
A Commercial Application of an Immobilized Enzyme,
High Fructose Syrup Production
Total Capital Investment . . . . . . . . . . . . . . . . . . 212
11.5
A Commercial Application of an Immobilized Enzyme,
High Fructose Syrup Production
Conversion Costs . . . . . . . . . . . . . . . . . . . . . . 213
12.1
Typical Nutrients for the Cultivation
of a Methylotropic Microorganism . . . . . . . . . . . . . . 221
Xiii