,.

10
4
M5

Authors : C . E . Rix
CONFIDE N TIAL October 23, 1968
C . N . Eaton
J . G . Jones Notebook Pages : 151501-151550,
158951-159000, 162501-162550,
16 3201-163250, 163301-163350,
163601-163650, 164301-164314,
Division : Chemical Rese arch 167901-167950, 168201-168222,
175551-175553 /6 sr 90/-/ 4a,7A s
RDR, 1968, No . 37 Dated : February 8, 1967 to
October 9, 1967
No . of Pages : 19 Previous Reports : None

XYLOSE PRODUCTION FROM CORN HULLS

~TND SUGAR L~AND 13~GASSE

OBJECT :

This report is concerned with optimizing the production of xylose-

rich syrup from corn hulls, a by-product of the corn wet milling industry,
and to a lesser extent with xylose production from sugar cane bagasse .
The xylose was to be used as a carbon source in the preparation of luq cose
isomerase, an enzyme which converts glucose to fructose .

SUMMARY :

In order to prepare xylose, which is used as a carbon source in

lucose isomerase production, destarched corn hulls and sugar cane bagasse
were ydro yzl ed with dilute sulfuric acid at temperatures from 100-150° C .,
with reaction times of 1 .0 to 4 .0 hr . The reaction mixtures were filtered,
the residue washed with water and the combined filtrates neutralized to
pH 4 .5 with calcium hydroxide, filtered, decolorized with charcoal and
deionized by passing through Amberlite IR-120 and Duolite A6 ion exchangers .
The deionized solutions were then concentrated under reduced pressure .
Corn hulls gave a 45% yield of a dry sugar syrup which contained approximately
30% arabinose, 52% xylose, 7% galactose, and 7% glucose for an overall
xylose yield of 23% . Sugar cane bagasse yielded 16% dry sugar syrup
containing 6% arabinose, 81% xylose and 9% glucose'for an overall xylose
yield of 13% .

However, in cell growth and lg ucose isomerase production, the xylose

samples, even after crystallization, had only 50-75% of the activity of
the xylose standard . Due to the low activity and the development of an
alkaline isomerization process for converting glucose to fructose this
project was terminated .
 2

TABLE OF CONTENTS
Page
OBJECT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

SUMMARY . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . 3

LIST OF FIGURES . . . . . . . . . . . . . . . . . . . . . . . . 3

A . INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . 4

B . EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . 4

I . Destarchi ng of Corn Hul l s . . . . . . . . . . . . . . 4

II . Acid Hydrolysis of Destarching Liquor . . . . . . . . 5

III . Hydrolysis of Corn Hulls Using Autoclave

Conditions . . . . . . . . . . . . . . . . . . . . . 6

IV . Pilot Plant Hydrolysis of Corn Hulls . . . . . . . . . 6

V . Atmospheric Hydrolysis of Sugar Cane Bagasse . . . . 6

VI . Atmospheric Hydrolysis of Corn Hulls . . . . . . . . 10

VII . Neutralization Conditions for Corn Hull

Hydrolyzate . . . . . . . . . . . . . . . . . . . . . 10
VIII . Optimizing Sugar Solution Decolorization
with Charcoal . . . . . . . . . . . . . . . . . . . . 10

IX . Amberlite IR-120 Ion Exchange Capacity . . . . . . . 13

X . Xylose Crystallization . . . . . . . . . . . . . . . 14

XI . Decomposition of Pure Xylose . . . . . . . . . . . . 14

C . DISCUSSION . . . . . . . . . . . . . . . . . . . . . . . . . 15

D . CONCLUSIONS . . . . . . . . . . . . . . . . . . . . . . . . 17

E . RECOMMENDATIONS . . . . . . . . . . . . . . . . . . . . . . 17

I . Future Work . . . . . . . . . . . . . . . . . . . . . . 17

II . Patentability . . . . . . . . . . . . . . . . . . . . . 17

BIBLIOGRAPHY . . . . . . . . . . . . . . . . . . . . . . . . . . 19
 3

LIST OF TABLE S

Table No . Title Page

I Destarching of Water-washed ,
Air-dried Corn Hulls . . . . . . . . . . . . 4
II Destarching of Wet, Sulfur Dioxid e
Containing Corn Hulls . . . . . . . . . . . 5

III Autoclave Hydrolysis of Destarche d

Corn Hulls with H2SO4 . . . . . . . . . . . 7
IV Pilot Plant Hydrolysis of Corn Hulls . . . . 9
V Atmospheric Hydrolyses : Corn Hull s
and Bagasse . . . . . . . . . . . . . . . . 9
VI Corn Hull Hydrolyzate Neutralizatio n
with Calcium Hydroxide . . . . . . . . . . . 10

VII Determining Color Absorbed per Uni t

Weight of Carbon (X/M)Co fo r
Nuchar C-190-N . . . . . . . . . . . . . . . 1 1
VIII Charcoal Efficiency in Suga r
Solution Decolorization . . . . . . . . . . 13
IX Amberlite IR-120 Depletion wit h
Calcium Hydroxide Neutralized Cor n
Hull Hydrolyzate . . . . . . . . . . . . . . 13

X Crystallization of Bagasse Xylos e

Syrups . . . . . . . . . . . . . . . . . . .14

XI Decomposition of Pure Xylose . . . . . . . . 14

LIST OF FIGURE S

Figure No . Title Page

1 Determination of (X/M)Co fo r
Nuchar C-190-N . . . . . . . . . . . . . . . 12
 4

A . INTRODUCTION

It had been found that xylose was the most desirable carbon source
for the production of lug cose isomerase, an enzyme that converts glucose
to fructose . Over the years, many agricultural by-products have served
as sources of x lose, some of which were : corn cobs (1-6), corn stalks (7),
oat hulls (6, 8~, cottonseed-hull bran (6, 9, 10), and sugar cane bagasse
(6, 9-11) . A promising material in this investigation appeared to be corn
hulls, a by-product of the corn wet-milling industry and readily available
from Penick & Ford . Analysis indicated that destarched corn hulls contained
38-41% pentosans . These pentosans are present in the hemicellulosic portion
of the hull and are in the form of a L-arabino-D-xyloglycan . The molecule
is primarily a xylan backbone with short side branches containing arabinose,
galactose, and D-glucuronic acid in a terminal position . Another xylose
source, sugar cane bagasse, is relatively inexpensive but the xylan content
is only about 20% . However, this was 80-90% xylose and purification was
greatly enhanced .

B . EXPERIMENTAL

I . Destarching of Corn Hulls

In a typical reaction, 100 g . of dried corn hulls (4 .4% H20, 8 .37%

starch, 38 .47% pentosan) was mixed with 2 .0 liters of water and heated at
reflux fo r 3 .0 hr ., filte red, washed with h ot water, a nd air dried .
Analysis : weight loss 19 .9%, starch 0 .98%, pentosan 40 .57% . Table I
and Table II give destarching conditions and results .

TABLE I

DESTARCHING OF WATER-WASHED,
AIR-D IED COWNHULLSy

Temperature Time % Starch % Pentosan Products of

Reaction °C . Hr . Wt . Loss, % Remaining Remaining Hydrolysisd
Untreated
Hulls - - - 8 .4 38 .5
1 100 1 .0 20 .8 1 .1 38 .8
2 100 2 .0 17 .6 0 .7 39 .3
3 100 3 .0 22 .9 0 .0 45 .3
4b 100 2 .0 19 .3 1 .3 52 .3
5 80-90 0 .5 8 .7 2 .1 39 .3 5% Arabinosee
21% Glucose
6 100 3 .0 18 .6 1 .3 39 .6
7 100 3 .0 19 .9 1 .0 40 .6
8c 25 1 .0 11 .8 7 .5 39 .0
a Liquid :hulls = 20 :1
b Ground to 20 mesh
c 2 .0 liters of 0 .25N H2SO4 :100 g . dry hulls
d Identifiable products in syrup from hydrolysis of destarching liquor
e Destarching liquor from 100 g . hulls yields 10 .5 g . of syrup on hydrolysis
 5

TABLE II

DESTARCHING OF WET, SULFUR DIOXIDE

CONTAIN VG CORN HUL SB- ~

Temperature Time % Starch % Pentosan Products ot

Reaction °C . Hr . % Wt . Loss Remaining Remaining Hydrolysis
Untreated
Hulls - - - 14 .9 31 .2
Pilot
Plant 1210 2 .0 - 1 .0 39 .8
Pilot
Plant 135° 1 .0 - 0 .1 33 .3 19% Xylose
34% Arabinose
Pilot
Plant 1000 2 .0 0 .5 38 .3 17% Maltose
36% Glucose
lc 80-90° 0 .5 19 .8 2 .4 38 .6 7% Arabinose
6% Glucose
2 100° 1 .0 10 .0 1 .0 37 .3
3 100° 2 .0 12 .5 0 .8 39 .3
4 100° 3 .0 24 .1 1 .0 39 :7
5 100° 4 .0 52 .4 1 .0 38 .7
6d 1000 2 .0 23 .3 1 .6 38 .1
7d 100° 1 .0 20 .6 2 .7 37 .8
8d 100° 1 .0 20 .3 2 .9 37 .5
9 100° 1 .0 21 .6 2 .2 39 .5
l0e 25° 1 .0 10 .1 12 .1 36 .0
lle 50-60° 1 .0 9 .6 11 .0 37 .4
a Liquid : hull ratio = 20 :1
b Identifiable products in syrup from hydrolysis of destarching liquor
c On hydrolysis of destarching liquor from 100 g . of hulls 12 .5 g . of
syrup obtained
d Ground to 20 mesh
e 2 .0 liters of 0 .25N H2SO4
:lOO g . dry hulls

II . Acid Hydrolysis of Destarching Liquor

Sulfur dioxide treated corn hulls were destarched, and an aliquot

taken of the destarching liquor . The solution was acidified to pH 2 .0
with con . H2SO4, refluxed for 6 .0 hr ., neutralized to pH 5 .0 with Ca(OH)2,
filtered, deionized with cationic Amberlite IR-120 and anionic Duolite
A-6 ion exchange resins and concentrated under reduced pressure . A 12 .5%
yield of brown syrup was obtained which contained 36% glucose and 17%
maltose . The results of other destarching liquor hydrolyses are presented
in Table I and Table II .
 6

III . Hydrolysis of Corn Hulls Using Autoclave Conditions

Destarched corn hulls (112 .8 g . wet, 100 g . dry basis) were placed
in a 4-1 . Erlenmeyer flask and acidified with 2000 ml . of 0 .12N H2SO4
(pH 1 .35) . The flask was placed in an autoclave set at 20 psi, 128° C .,
heated for 20 min . to reach equilibrium, and then heated for an additional
1 .0 hr . The mixture was filtered, and the residue slurried with 500 ml .
of water, filtered, and the filtrates combined . The combined hydrolyzate
solution was neutralized to pH 4 .5 with calcium hydroxide, boiled for
15 min . and filtered hot . The solution was then passed through a 400 ml .
granular charcoal column, 500 ml . of Amberlite IR-120, 500 ml . of Duolite
A-6 and the final pH was adjusted to 5 .0-5 .5 with small amounts of IR-120 .
The solution was concentrated under reduced pressure at 60° C . to yield
52 .3 g . of dry syrup containing 33-34% arabinose, 54% xylose, and 9-10%
glucose and galactose . The total yield of xylose (based on weight of dry
hulls) was 28 .2% . Various reaction conditions and yields are presented
in Table III .

IV . Pilot Plant Hydrolysis of Corn Hulls

In the pilot plant studies a horizontal, rotating, stainless steel

reactor coated internally with polyvinyl chloride was used . The cylindrical
reactor was steam jacketed and possessed internal baffles . The steam
pressure and internal temperature could be measured directly . The reactor
was charged with 2 .27 kg . (5 .0 lb .) of dry destarched corn hulls and
27 .3 kg . of 0 .1 N H2S04 and was heated to 148° C . (50 psi . steam pressure)
and maintained at this temperature for an additional 0 .5 hr . The reaction
mixture was centrifuged, and the filter cake washed with 6 .0 liters of
water, yielding 29 .8 kg . of hydrolyzate and 1 .15 kg . of dry residue (7%
pentosan remaining) . A 1 .32 kg . aliquot of the hydrolyzate was neutralized
to pH 4 .5 with calcium hydroxide, boiled 0 .25 hr . and filtered hot . The
filtrate was passed through 400 ml . of granular charcoal, 500 ml . of
Amberlite IR-120, 500 ml . of Duolite A-6, and the final pH was adjusted
to ti5 .0 with small amounts of Amberlite IR-120 . On evaporation at 60° C .
and reduced pressure, 39 .4 g . of dry syrup was obtained . Analysis showed
26% arabinose, 55% xylose, 7% galactose and 8% glucose for a total yield
of xylose based on dry hulls of 21 .5% . Various reaction conditions and
yields are presented in Table IV .

V . Atmospheric Hydrolysis of Sugar Cane Bagasse

Sugar cane bagasse (460 g . wet, 200 g . dry basis) was placed in a
5 liter round bottomed flask containing 2000 g . of 3 .75% H2SO4 and heated
at reflux with stirring for 3 .0 hr . The purification technique was as
above, yielding 33 .1 g . of dry syrup containing 6% arabinose, 81% xylose,
2% galactose, and 7% glucose . The total yield of xylose, based on dry
weight of bagasse,was 13 .2% . Various reaction conditions and yields are
presented in Table V .
 TABLE III

AUTOCLAVE HYDROLYSIS OF DESTARCHED CORN HULLS WITH H2SO4

Experiment Liauid Acid % Dry % Dry % % % Glucose Total %

No . Hulls Conc . N Time/Hr . Residue Syrup Arabinose Xylose and Galactose Xylose

163302 20 :1 0 .07 2 .0 45 23 .5b 18 60 12 14 .1

163303 " 0 .12 2 .0 41 15 .7b 17 50 11 7 .9
163304 " 0 .03 3 .0 46 32 .9b 20 50 11 16 .4
163305 " 0 .07 3 .0 42 39 .Ob 18 48 13 18 .6
163306 " 0 .12 3 .0 43 23 .0b 23 64 16 14 .6
163307 " 0 .03 4 .0 53 19 .0b 19 52 13 9 .8
163308 " 0 .07 4 .0 45 47 .7b 21 59 17 28 .4
163309 " 0 .12 4 .0 42 .1 22 57 17 24 .0
163311a " 0 .07 3 .0 42 32 .3 20 40 8 12 .9
163312a " 0 .12 3 .0 44 31 .6 18 42 10 13 .3
163314a " 0 .03 4 .0 51 23 .4 19 25 6 5 .9
163315a " 0 .07 4 .0 45 36 .8 19 40 9 14 .7
163316a " 0 .12 4 .0 53 30 .6 19 47 9 14 .4
163317 " 0 .07 4 .0 47 26 .6 14 62 22 16 .5
163320 " 0 .07 4 .0 40 32 .0 18 49 14 16 .0
163323 " 0 .20 2 .0 41 43 .4 23 60 15 26 .0
163324 " 0 .30 2 .0 42 30 .8 20 51 16 15 .8
163325 " 0 .40 2 .0 40 20 .9 21 56 18 11 .7
163326 " 0 .20 1 .0 33 39 .6 31 54 13 21 .5
163327 " 0 .30 1 .0 42 32 .6 29 52 13 17 .0
163328 " 0 .40 1 .0 48 46.5 27 49 14 22 .0
163329 3 :1 0 .20 2 .0 37 15 .0 26 46 7 6 .9
163330 3 :1 0 .30 2 .0 37 6 .5 25 44 12 2 .8
163331 3 :1 0 .40 2 .0 50 50 .0 25 44 8 2 .2
163334 20 :1 0 .20 1 .5 33 30 .4 27 47 11 14 .4
163335 " 0 .30 1 .5 34 45 .7 29 51 12 23 .3
163336 " 0 .40 1 .5 29 47 .7 31 54 14 25 .8
163337 " 0 .12 1 .0 34 43 .4 30 55 10 23 .8
163338 " 0 .16 1 .0 27 22 .8 32 59 13 13 .4
163339 " 0 .25 1 .0 28 23 .4 30 54 9 12 .7
163340 " 0 .20 2 .0 37 45 .9 27-31 47-54 8-13 21 .6-24 .8

hbi6 96009
 TABLE III Cont'd

AUTOCLAVE HYDROLYSIS OF DESTARCHED CORN HULLS WITH H2SO4

Experiment Liquid Acid % Dry % Dry % % % Glucose Total %

No . I-1 ,j 11 S Conc . ,f~ Ti,melNr, Rasi,d u o $ ru n Ar a hi,nogo Xylose Galactose X ~y,ose
163341 20 :1 0 .20 2 .0 30 51 .4 29 57 14 28 .6
m
163342 0 .20 2 .0 34 36 .7 29 55 13 20 .0
163345 m 0 .12 1 .0 33 52 .4 33-34 54 9-10 28 .2
163346 0 .20 1 .0 37 43 .7 31-33 50-53 11-12 22 .0
163347 0 .30 1 .0 32 39 .8 34 55 12 22 .0
167901 0 .20 2 .0 37 44 .2 34 57 13 25 .0
167902 0 .20 2 .0 32 43 .2 32 56 14 24 .1
167903 0 .20 2 .0 37 43 .8 32 52 13 22 .8
167908 3 :1 0 .20 4 .0 38 38.8 30 52 14 20 .6
167909 3 :1 0 .40 4 .0 38 38 .4 30 51 16 19 .6
167910 20 :1 0 .20 4.0 32 46 .0 31 51 15 23 .4
167911 20 :1 0 .40 4.0 33 46.5 29 46 17 21 .4

aH3P04 used

bwet

9bi6 96009
 TABLE IV

PILOT PLANT HYDROLYSIS OF CORN HULLS

Experiment Liquid : H2SO4 % % Dry % % % Glucose Total %

N uM lber Hul l s Conc . N T i:^e/Hr . Temn
r • °C . Residue Syrup Arahi-se Xylose and Galactose Xylose
163343 10 :1 0 .40 1 .5 128 40 35 .7 35 47 8 16 .8
167904 10 :1 0 .20 1 .33 142 35 39 .4 31 47 13 18 .5
167906 10 :1 0 .20 1 .5 147 32 36 .6 29 42 13 15 .4
167919 3 :1 0 .10 0 .5 126 61 17 .3 45 35 0 6 .1
167938 12 :1 0 .10 0 .5 148 49 39 .1 26 55 15 21 .5
167921 3 :1 0 .30 0 .5 148 42 20 .0 29 51 8 10 .2
167945 12 :1 0 .30 0 .5 126 34 35 .9 30 46 21 16 .5
167924 3 :1 0 .30 3 .0 148 39 15 .5 26 35 14 5 .4
167925 12 .1 0 .30 3 .0 126 28 35 .6 29 48 8 17 .1
167926 3 :1 0 .30 3 .0 126 38 34 .9 26 48 7 16 .7
167927 12 :1 0 .30 3 .0 148 36 22 .6 24 28 15 6 .4
167929 3 :1 0 .10 3 .0 148 34 9 .1 29 46 14 4 .2
167930 12 :1 0 .10 3 .0 126 42 37 .6 30 52 14 19 .6
167932 7 .5 :1 0 .20 1 .75 138 39 33 .5 27 48 14 16 .0
167933 7 .5 :1 0 .20 1 .75 138 36 32 .6 27 48 12 15 .6

TABLE V

ATMOSPHERIC HYDROLYSES : CORN HULLS AND BAGASSE

H2SO4 % % Dry % % % Glucose Total %
Material Liquid :Hulls Conc . % Time/Hr . Residue Syrup Arabinose Xylose and Galactose X ly ose
Hulls 10 :1 4.2 3 .25 38 19 .4 30 50 12 9 .7
Hulls 7 .5 :1 5 .6 2 .00 36 45 .2 26 51 11 23 .1
Bagasse 10 :1 3 .75 3 .00 78 16 .6 6 81 9 13 .2
Bagasse 12 :1 4 .0 3 .00 74 11 .8 2 90 4 10 .6
Bagasse 16.7 :1 4 .4 0 .83 77 12 .3 3 87 4 10 .7

9hi6 96009
 10

VI . Atmospheric Hydrolysis of Corn Hulls

In a 5 liter round-bottomed flask were placed 453 .5 g . of wet corn

hulls (11 .8% H20, 400 g . dry basis) and 3000 g . of 5 .6% H2SO4 . After 0 .5
hr . the mixture had reached 100° C . and was then refluxed with stirring
for 2 .0 hr . The purification technique was as above, yielding 181 .2 g .
of dry syrup containing 26% arabinose, 51% xylose, 6% galactose, and 5%
glucose . The total yield of xylose, based on dry weight of hulls, was
23 .1% . Various reaction conditions and yields are given in Table V .

VII . Neutralization Conditions for Corn Hull Hydrolyzate

The hydrolyzate had an initial pH of 1 .22 . The transmittance (T)
equaled 75 .5% at a wavelength (x) of 520 mu . The theoretical sulfate
concentration was 0 .79% . Calcium hydroxide was added to 100 ml . aliquots,
the pH change measured, the solution boiled 15 minutes and filtered hot .
The resulting pH, transmittance, [Ca+2], and [SO4-2] were measured and
it was found that a pH range of 3 .5 to 4 .5 optimized clarity and sulfate
ion removal without undue buildup of calcium . The results are recorded
in Table VI .

TABLE VI

CORN HULL HYDROLYZATE NEUTRALIZATION

WITH CALCIUM HYDROXIDE

Wt . Ca(OH)2 % T
pH Added a=520 mu [Ca+2] M . [SO4-2] M .
1 .23 0 .0 75 0 .000 0 .082
2 .95 0 .555 g . 61 0 .021 0 .028
4 .15 0 .618 72 0 .025 0 .025
4 .92 0 .669 64 0 .027 0 .029
5 .90 0 .687 56 0 .031 0 .022

VIII . Optimizing Sugar Solution Decolorization with Charcoal

A neutralized (pH 4 .5) corn hull hydrolyzate with initial color

(Co) = 0 .95 at 420 mu as measured by a Bauch and Lomb Spectronic 20
was used in all experiments . The pulverized carbon to be tested was
added in increasingly larger dosages (M) to five 100-ml . portions of
sugar solution, stirred for 1 .0 hr . and filtered twice . The residual
color (C) was measured, and the color absorbed (X = Co -C) was calculated .
The color absorbed per unit weight of carbon (X/M) was also calculated and
plotted versus residual color (C) on log-log paper . If a vertical line
is erected from the point on the horizontal scale corresponding to the
influent concentration (Co), and the isotherm is extrapolated to intersect
that line, the (X/M) value at the point of intersection can be read from
the vertical scale . This value, termed (X/M)co, represents the amount
of impurity absorbed per unit weight of carbon when that carbon is in
 11

equilibrium with the influent concentration . It represents the ultimate

capacity of the carbon and is used as the standard of comparison . Typical
data for (X/M)Co calculations are recorded in Table VII and Figure 1 .
The comparison of charcoal decolorizing efficiencies shown in Table VIII
indicates that Nuchar C-190-N is the most effective decolorizing agent
both at 23° C . and 50° C .

TABLE VII

DETERMINING COLOR ABSORBED PER UNIT WEIGHT

OF CARBON X/M)Co FOR NUCHAR C-190-N

Color Absorbed
Initial Residual Color Absorbed per Unit Wt .
Wt . Carbon (M Color C) Color C CO-C = X of Carbon (X/M)
0 .95
.1 .55 .40 4 .00
.2 .34 .61 3 .05
.3 .21 .74 2 .47
.4 .14 .81 2 .02
.5 .07 .88 1 .76
(X/M)co = 5 .1
 12

FIGURE 1

DETERMINATION OF (X/M)CO FOR NUCHAR C-190-N

~~ i--
7

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I'
• i.

: _ _
` „. , .,, .
-
..
EIIEIL
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_; ~ : -.
.; :I; :. . .
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7-77 - -: ;

=
. , . .. : . . . . . . . . . ~ +
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~, , ,
l . . . ~ ~-
t
~

• -, . . ., .~ _ :. : .. . . . }y r ~i ~ ; ,, i .,
t
; ~,1 ~y ,{a r ,+r }, f~, i4
~ ; .i - ~ ;y i ,{+ 4 - Y+ . .
; ~ T]
, 1 #il} I,
,, n'
>1 c2 .03 o4 o5 .c+& .07 .c8A .10 .2 .3 .4 .5 1 .6 .1 .8 .9 ~
x
RESIDUAL SOLUTION COLOR (C)
(D

11
 13

TABLE VIII

CHARCOAL EFFICIENCY IN SUGAR SOLUTION DECOLORIZATION

Carbon Sample (X/M)co 23° C . (X/M)co 500 C .
Duo S-51 4 .3 5 .7
Nuchar CE :E-N 3 .5 4 .1
" C-190-N 5 .1 6 .0
" C-115-N 3 .83 4 .30
" Aqua A 3 .0 2 .46
" CE:E A 4 .9 -
Pittsburg CAL pulverized 3 .88 3 .90
" CPG pulverized 3 .10 3 .20
" SGL pulverized 3 .0 2 .05

IX . Amberlite IR-120 Ion Exchange Capacity

A 500 ml . column of wet Amberlite IR-120 was regenerated with 600 ml .

of 3N HC1 and exhausted with a CaSO4 solution from a neutralized corn hull
hydrolyzate . The results in Table IX indicate effective ion removal from
a 0 .035 M . Ca+2 solution whose volume was sixteen times greater than the
wet resin bed .

TABLE IX

AMBERLITE IR-120 DEPLETIONa WITH CALCIUM

HYD I EIITWIZED COlt~}TULT~RI'DT'btYZA'f E b
Volume
Collected 0 ml . 500 3300 8000 9000 11,500 13,000
pH
Effluent 2 .0 2 .0 2 .0 2 .2 2 .3 4 .3 4 .4
[Cr]mg ./1 . 0 .2 0 .08 0 .25 0 .16 0 .20 17 .25 20 .84
Ca 1384 .0 0 .36 0 .29 7 .70 293 .9 564 .0 1054 .0
Fe 2 .67 0 .41 0 .52 2 .70 12 .12 20 .2 52 .2
Ni 0 .07 0 .10 <0 .1 0 .10 1 .43 6 .0 10 .25
Cu 0 .11 0 .02 <0 .02 <0 .02 0 .09 0 .02 0 .03
K 31 .73 0 .05 3 .53 3 .50 4 .22 97 .3 42 .3
Na 39 .18 4 .95 4 .66 5 .5 11 .20 195 .0 30 .0
Zn 9 .43 <0 .03 - 0 .42 11 .56 26 .3 20 .45
Mn 39 .52 <0 .02 - 2 .50 34 .40 70 .8 67 .90
Mg 7 .57 0 .49 0 .11 12 .00 86 .70 183 .0 158 .9
a A 500 ml . column of wet resin regenerated with 600 ml . of 3N HC1 .
b From stainless steel reactor .
 14

X . Xylose Crystallization

Because of the larger xylose content, syrups from sugar cane bagasse
showed a greater tendency to crystallize than those syrups obtained from
corn hulls . The samples (5% arabinose, 77% xylose, and 8% glucose) were
heated to dissolve the sugar and cooled slowly with shaking to 24° C .
Syrups containing 80% solids yielded 63% crystalline material . See
Table X .

TABLE X
CRYSTALLIZATION OF BAGASSE XYLOSE SYRUPSa

Initial, % Solids After Cr stallizationb

~ ~ % o i ater a
<70 100 Trace of fine crystals
70 82 .2 17 .8
75 55 .7 44 .3
80 36 .7 63 .3
85 Solidification rather than crystallization

a 5% arabinose, 77% xylose, 8% glucose

b Sugar analysis on crystalline material 0% arabinose, 96% xylose, 2% glucose

XI . Decomposition of Pure Xylose

A 1% xylose solution in 0 .20 N H2SO4 was heated for 4 .0 hr . at 128° C .

(20 psi steam pressure) . Ultraviolet analysis and gas chromatography
showed the formation of furfural, crotonaldehyde and a small amount of
water-insoluble brown polymer . Table XI gives a breakdown of the decom-
position products .

TABLE XI
DECOMPOSITION OF PURE XYLOSE

% of
Starting Material
Initial wt : . xylose 1 .877 g .
Xylose as furfural in condensate 0 .052 g .
Xylose as furfural and crotonaldehyde `~ 10 .5%
in solution 0 .145 g . _
Unidentified polysaccharide .32 g . 17 .0%
Recovered xylose 1 .300 69 .5%
Polymeric material trace

97 .0%
 15

C . DISCUSSION

In the production of xylose from corn hulls, three general steps

are involved : (1) removing residual starch from the corn hull, (2) acid
hydrol sis of the destarched hull to produce a dilute sugar solution,
and (3~ purification of the hydrolyzate to yield an acceptable xylose
product . Xylose production from bagasse is similar ; however, the initial
destarching is unnecessary .

Any residual starch on the hulls will be hydrolyzed to glucose, and

the glucose in turn will inhibit lucose isomerase production . The
starch may be removed from the hulls by hot water solubilization . The
initial starch content of 8-15% was reduced to 1% by refluxing an aqueous
suspension of the hulls for 3 .0 hr ., or heating at 121° C . (15 psi . steam
pressure) for 2 .0 hr . See Table I and Table II . In addition to starch
removal, there was a weight loss of approximately 20% due to dissolution
of plant gums . Cold dilute acid treatments appeared less efficacious
for starch removal than boiling water .

When the corn hulls were heated with dilute sulfuric acid, the
hemicellulosic xylan chains were broken down in preference to the glucose
links in the cellulose . Xylose and arabinose were the principal products
of mild hydrolysis, though galactose and glucose were also found . If
reaction conditions became too severe the xylose formed initially was
decomposed to furfural, crotonaldehyde, and polymeric material . See
Table XI .

On a laboratory scale a 7 .5 :1 ratio of 5 .6% sulfuric acid to dry,

destarched hulls heated at reflux for 2 .0 hr . produced a 45% yield of dry
sugar syrup . The syrup contained 26% arabinose, 51% xylose, and 11%
galactose and glucose for an overall xylose yield of 23% . See Table V .
Using an autoclave set a 20 psi . steam pressure (128° C .), overall yields
of xylose as high as 28% have been attained . See Table III .

In the pilot plant, reactions were carried out in a horizontal,

polyvinyl chloride coated reactor . On treating 2 .27 kg . (5 .0 lbs .) of
dry hulls with 27 .3 kg . (60 lbs .) of 0 .1 N H2SO4 and heating at 148° C .
for 0 .5 hr ., a 21 .5% overall yield of xylose was obtained . This accounts
for 90-95% of the xylose initially present in the hull . Statistical
analysis performed on the data from pilot plant reactions (See Table IV)
have produced equations for xylose yield estimation . One polynomial
having all first degree terms is given below . On filling in values for
the variables an estimate of the total percent yield of xylose will be given .
X10 = .76X1 - 3 .39X2 - .85X3 - .lOX4 + 38 .7 (See 5 .4)
where X1 3 :1 < ratio liquid to hulls < 12 :1

X2 0 .1N < [H2SO4] < 0 .3N

X3 0 .5 hr . < time < 3 .0 hr .
 16

X4 258° F . < temperature < 298° F .

X10 0% < total % xylose < 28%

The term See 5 .4 indicates that the answer given will be within ± 5 .4%
absolute value of the predicted yield 65% of the time . In the variable
ranges given above an increase in liquid to hull ratio should produce
larger yields, as would a temperature decrease . Reaction time has very
little effect on yield and acid strength none at all .

After the various sugars in the corn hulls or sugar cane bagasse
had been liberated by acid hydrolysis various purification steps were
necessary . The degree of purity determined thoroughness of the reaction
workup .

To neutralize the solution and remove sulfate ions, the pH was adjusted
to 4 .5 with calcium hydroxide, boiled 15 minutes and filtered . The decreased
solubility of calcium sulfate at high temperature permitted removal of
almost all of the sulfate ions, leaving only a small amount of calcium
sulfate in solution ([ CaSO4] = .025M ti3 .0 g ./l . - see Table VI) .

The neutralized sugar solution was then passed through a column of

granular charcoal to remove color bodies and high molecular weight impurities
which caused severe frothing when vacuum evaporation of the solution was
attempted . The water white effluent had a pH of approximately 6 .0 after
passing through charcoal .

The last traces of ionic m~terial were removed with ion exchange
resins . Cations, primarily Ca+ , were removed with Amberlite IR-120 .
The resin was quite effective and total metal ion concentration was reduced
to less than 10 mg ./1 . for a 2% xylose solution (See Table IX) . At this
low ionic concentration Na+ and K+ were the most prevalent species . As
the resin approached exhaustion, loosely held ions were displaced by
incoming polyvalent species . This undesirable situation was carefully
avoided in sample preparation . Anions, primarily S04=, and some color
bodies were removed with an anionic Duolite A-6 resin . The effluent
showed no S04= leakage when tested with BaC12 . Ash analysis indicated
no inorganic material was present .

The bagasse syrup (5% arabinose, 77% xylose, 8% glucose) containing

a higher percentage of xylose than the corn hull syrup (33% arabinose,
54% xylose, 9% glucose and galactose) crystallized much more readily and
afforded a method of purification . Upon crystallization, bagasse syrups
yielded a product containing 96% xylose and 2% glucose . However, 30%
of the total xylose remained in the mother liquor (See Table X) . Syrups
obtained from corn hulls will crystallize but usually require several
weeks to do so .

After the xylose syrups had been obtained, they were submitted for
biological testing, and their ability to produce glucose isomerase was
measured against a standard xylose solution . Crysta ize samp es containing
 17

96-98% xylose showed a cell growth activity at best only 75% of that of
the standard . Chemical analyses were not able to isolate any trace materials
responsible for the difference in cell growth activity .

D . CONCLUSIONS

The corn hulls, readily available from the corn wet milling industry,
have a pentosan content of approximately 40% . The pentosans present as
arabinose and xylose were liberated by acid hydrolysis and purification
yielded colorless syrups containing 30% arabinose, 52% xylose, 7% galactose
and 7% glucose . However, the xylose in the syrup was only 45% as effective
as the standard Eastern Chemical xylose for glucose isomerase production .
Although xylose rich syrups were produced easily and n ood yields, the
additional arabinose, glucose and trace materials present markedly decreased
their ability to function as a carbohydrate source for g~l_~u~~_c~ose isomerase
production . Sugar cane bagasse contains ti20% pentosan, a- n~fc upon ac~i3~
hydrolysis yielded water white syrups containing 6% arabinose, 81% xylose,
2% galactose, and 7% glucose . On crystallization, a product containing
96% xylose was obtained . Even this product, however, was only 50-75%
as effective as Eastern Chemical Company xylose in the production of
9-lu-co-se- isomerase . This lessened cell growth efficiency coupled with the
advent of an economical alkaline isomerization process for converting
glucose to fructose led to termination of this project .

E . RECOMMENDATIONS

I . Future Work

At this time no further work is anticipated .

II . Patentability

The production of xylose from corn hulls has not been previously
reported in the literature . However, the general methods employed are
similar to those used in xylose production from other hemicellulosic
materials . Therefore, acid hydrolysis of corn hulls to yield xylose
would probably not be patentable .

~ aton

~
- .-V ~ _____J 0
. G . Jones
Approved :

(See next page for distribution .

 18

Distribution :

Dr . Murray Senkus Mr . M . R . Haxton

Dr . R . E . Farrar Mr . L . A . Willson, Jr .
Mr . E . H . Harwood Dr . H . J . Bluhm
Dr . C . E . Teague, Jr .
Dr . C . E . Rix
Miss C . N . Eaton
Mr . J . G . Jones
,Xibrary (2)
Dr . Edward Bernasek

Submitted : October 23, 1968

Completed : October 25, 1968

From manuscript :bjv ;kti
 19

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1. Hudson, C . S ., and Harding, T . S ., J . AM . CHEM . SOC . 39, 1038 (1917) .

2. Hudson, C . S ., and Harding, T . S ., Ibid . 40, 1601 (1918) .

3. Monroe, K . P ., Ibid . 41, 1023 (1919) .

4. Ling, A . R ., and Nanji, D . R ., J . CHEM . SOC ., 123 (pt . 1) 620 (1923) .

5. Whistler, R . L ., and Wolfrom, M . L ., "Methods in Carbohydrate
Chemistry," Vol . 1, Academic Press, New York, New York, p . 88 (1962) .
6. Dunning, J . W ., and Lathrop, E . C ., IND . ENG . CHEM . 37, 24 (1945) .

7. Firstenberger, B . G ., Iowa State Jour . Sci ., 18, 27 (1943) .

8. Bryner, L . C ., Christensen, L . M ., and Fulmer, E . I ., IND . ENG . CHEM .

28, 206 (1936) .

9. Schreiber, W . T ., Geib, N . Y ., Wingfield, B ., and Acree, S . F .,

Ibid . 22, 497 (1930) .

10 . Scherrard, E . C ., and Blanco, G . W ., Ibid ., 12, 1160 (1920) .

11 . Ledoga S . p . A ., Brit . Pat . 922,684, April 3, 1963 .

12 . Whistler, R . L ., Ed ., "Industrial Gums," Academic Press, New York,

New York, p . 301 (1959) .