BIOTECHNOLOGY-lecture note-PART7

Bio-Chemical conversion

Biochemical conversion of biomass is defined as the converting of biomass raw materials into gaseous
or liquid fuels, such as biogas or bioethanol, using bacteria, micro-organisms and enzymes. Anaerobic
digestion (AD) and fermentation are the major biochemical technologies. In following sections, both
biochemical technologies will be scrutinized. Other biochemical conversion are being studied and
developed such as hydrogen from some specific types of organism living in an aqueous environment.

3.1 Anaerobic digestion (AD)

Anaerobic digestion (AD) is a process in which biomass raw materials directly converted to bio-gas
by anaerobic microorganisms. The bio-gas is the combination of methane (more than 50%wt), carbon
dioxide (CO2), carbon monoxide (CO), nitrogen (N2), hydrogen (H2), and in small quantities hydrogen
sulphide (H2S). The bio-gas can be handled in CHP-generating stations to generate electricity and
thermal energy or, energy carriers after further processing, upgrading, and transportation to be applied
as an alternative to fossil fuels as well. The produced bio-gas has 20-40% of feedstock heating value.
Sewage sludge, agricultural waste, animal waste, food waste, and industrial waste (e.g., bio-solids,
organic wastewater, etc.) as the high-moisture organic wastes are appropriate feedstocks for AD
process by overall conversion efficiency of 21%. Produced bio-gas can be directly utilized in spark-
ignition gas engines and gas turbines. It can be also upgraded to higher quality natural gas by removing
carbon dioxide. Figure illustrates the feedstocks, anaerobic digestion system, and its products.

Figure 1: Feedstocks, system, and products of Anaerobic digestion.

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Anaerobic digestion process
As mentioned before, the AD can chemically breakdown the feedstocks to produce bio-gas. The
breaking-down reactions are scrutinized in several stages as shown in Figure .

Figure 2: Schematics of AD steps

There are four main steps of AD, containing hydrolysis, acidogenesis, acetogenesis, and
methanogenesis. Each step handles an individual types of microorganisms to metabolize material.
The first step is hydrolysis in which bacteria decompose fat, protein, and carbohydrate and convert
them into soluble organic molecules such as sugars, amino acids, fatty acids, alcohol, carbon dioxide,
ammonia, hydrogen, and sulphides.

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 In the second step (i.e., acidogenesis), microorganisms decompose the yields of the first (i.e.,
hydrolysis) step to produce hydrogen, carbon dioxide, acetic acids, and volatile fatty as well as some
alcohols.
At third step, the volatile fatty acids are decomposed and produce carbon dioxide, hydrogen, water
and acetic acid during acetogenesis process.
Finally in the fourth step, Methanogenic bacteria can produce the methane (CH4) as a bio-gas,
carbon dioxide (CO2) and water (H2O).

Kinds of Digesters

1- Covered Lagoon Digester

Cover of lagoon is constructed by an impermeable plastic membrane to solve the odor problems which
spread during the break down process of the manure. This model of digester is applied in warm times.
Figure illustrates one Covered lagoon digester and its internal structure.

Figure 3: Covered Lagoon Digester and its internal structure

2- Complete Mix Digester

This kind of digester is applied in digestion of manure containing 2-10 percent solids. A gas-mixing
system keeps the solid fractions in suspension and reinforces the rate of digestion process in heated
tank. These digesters are more expensive than the covered lagoon digester.

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 Figure 4: Complete Mix Digester and its internal structure

3- Plug-Flow Digester
Plug-flow digesters are suitable for the converting of animal manure, including a solid concentration
of 11-14 percent. In this kind of digester, the solids in manure partially settle in bottom of the tank.
Since this digester has few motive parts, it requires less maintenance.

Figure 5: Plug-Flow Digester and its internal structure

4- Temperature-Phased Anaerobic Digestion (TPAD)

The Temperature-Phased Anaerobic Digester (TPAD), is the combination of the conventional
digestion methods in a two-step reactor. The first step is processed at high temperature (i.e., 55℃), and
the second step runs at lower temperature (i.e., 35℃). Utilization of more dilute manures of dairy farms
and heating process enhance the performance of mentioned digester.

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 Figure 6: schematic of TPAD system.

5- Effective parameters on the AD

The monitoring and controlling of the physical and biochemical conditions during the process is
essential to conserve the biological population and improve the production and quality of biogas. These
physical and biochemical parameters include temperature, pH, alkalinity, nutrients and trace elements,
total solids and volatile solids, organic loading rate, and retention time.

3.2 Aerobic Fermentation

The aerobic fermentation biochemically converts the biomass feedstocks into bioethanol by normal
metabolism in the presence of oxygen conducted in yeasts. In this method, sugar, starch, and cellulose
are metabolized by cells during the fermentation process to produce the bioethanol as one of the
important fuels all over the world with 106 billion liters production in 2017 [126]. Bio-ethanol can be
used as pure alcohol or blended with conventional gasoline to fuel engines and/or turbines after
upgrading process (e.g., distillation and dehydration). [2]. The overall chemical reaction of
fermentation is presented in equation of (11).
C6H12O6 → 2C2H5OH+2CO2 (11)
In comparing to AD, fermentation is shorter and more favorable process but oxygen limitation is
an important disadvantage of fermentation, because oxygen almost cannot be dissolved in water.

Feedstocks and processes

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 Figure 7: General scheme for the feedstocks of aerobic fermentation

According to Figure 7, miscellaneous biomass resources of biomass can be appropriate for aerobic
fermentation technology of conversion. Among these raw materials, lignocellulosic biomass with
carbohydrates building block have the most sustainability and the lowest energy content. On the other
hand, the edible oil-crops and corn with the highest fatty acids structures, have the maximum energy
content and cost. Many microorganisms, including bacteria and yeasts, can be used as biocatalysts and
can accelerate the fermentation process. It should be noted, the energy content of foods with high fatty
acids is higher than foods with higher hydrocarbons as illustrated in Figure 7.
In commercial-scale fermentation, yeasts are more suitable than bacteria, because of having some
positive factors which leads to higher amount of ethanol production. These factors include the
thickness of their cell walls, big sizes, higher resistance to contamination, negligible nutritional
requirements, and better growth at acidic pH. Saccharomyces cerevisiae, Pichia stipites, and Candida
stipites are the most promising yeasts in microbiological fermentation. Also, Escherchia coli and
Zymomonas are the famous kinds of bacteria.

Fermentation of first generation biofuel feedstocks

According to Figure 8, first generation bioethanol is produced from sugar (sucrose-containing
feedstocks) and starchy biomass after several proper pre-processing, fermentation, and post-processing
steps [129].

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 Figure 8: Feedstocks, fermentation, and upgrading steps in sugar and starchy biomasses

Sugar crops fermentation

Sugar crops (e.g., sugar beet, sugar cane and sweet sorghum) mostly are comprised by glucose,
fructose, and sucrose [130]. The mentioned fermentable sugar crops are pre-processed by milling or
crushing before the fermentation step. During fermentation, yeast converts sugars into alcohol and
CO2. After fermentation, resulted bio-ethanol is purified by distillation followed by dehydration after
fermentation as depicted in Figure .

Figure 9: Steps of sugar cane fermentation schematically.

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Starchy (Grains) crops fermentation
Starchy crops such as corn and wheat contain starch, which is a polysaccharide of glucose units linked
by α (1-4) and α (1-6) glycosidic bonds. Starch is not directly fermented by yeast. After milling the
raw materials and extracting starch, the obtained starch is hydrolyzed and converted into glucose using
α-amylase and glucoamylase.. The produced glucose is then fermented to ethanol.
The milling process of the starch is conducted by one of both methods of dry grinding or wet milling
process [112]. The main discrepancy between these two processes is the obtaining procedure of
glucose and co-products [113]. In dry grind, whole corn is milled to produce ethanol accompanied to
high protein animal feed called dry distillers’ grains with soluble (DDGS). During wet milling, soaking
of corn is followed by separation of germ, fiber, and starch. Wet milling produces some by-products
such as corn sweeteners, oil, and corn gluten meal beside the ethanol. Wet milling is most expensive
and less efficient in producing ethanol than dry grinding process [129].

Figure 10: Schematics of pretreatment, fermentation, and upgrading processes of starchy

feedstocks.

Fermentation of second generation biofuel feedstocks

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 Figure 11: Feedstocks, fermentation, and upgrading steps in sugar and starchy biomasses

Since mentioned before, the lignocellulosic raw materials (e.g., woods and straw) are known as the
second generation feedstocks. Lignocellulosic biomass can be altered into glucose and fundamentally
xylose by the use of enzymatic hydrolysis. Figure illustrates the main stages of the ethanol production
from the lignocellulosic feedstocks.

Figure 12: Main steps of cellulose fermentation.

According to Figure 12, pre-processing and pre-treatment process is conducted before the
fermentation process to destroy the cell wall and provide the cellulose, hemicellulose fibers for further
processing. Pre-treatment methods are mainly categorized into four groups comprising:
(i) Physical (milling and grinding),
(ii) Physiochemical (steam pretreatment/auto hydrolysis, hydro-thermolysis, and wet oxidation),
(iii) Chemical (alkali, dilute acid, oxidizing agents, and organic solvents),
(iv) Biological or a combination of these methods [128].
These pre-processing approaches can decompose the polymeric units to produce pentose sugar (i.e.,
monosaccharide with five-carbon atoms) from cellulose and hexose sugar (i.e., monosaccharide with
six-carbon atoms) from hemicellulose for the fermentation process. Monosaccharides cannot be
hydrolyzed to smaller carbohydrates.