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Industrial production of lactic acid and its applications

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Review Article: IJBR
Dec 2018: Volume 1, Issue 1

Industrial production of lactic acid and its applications

Battula Savithra Krishna*, Gantala Sarva Sai Nikhilesh1, Besetty Tarun1,

Narayana Saibaba K V1, R Gopinadh1

Department of Biotechnology, GITAM Institute of Technology, GITAM University

(Received 20 December, 2018 Accepted 26, 2018)
Corresponding author E-mail address: bskrishna1999@gmail.com
Key words: Lactic acid, Solid State Fermentation, Bacteria, Fungi, Renewable raw materials.
Abstract

Lactic acid is one of the most commercially useful hydroxycarboxylic acids. Its applications
range from bulk production of products, like Poly Lactic Acid (PLA), in industries to simple
house hold applications such as food containers. Many cheap materials, such as starchy and
cellulosic materials, and renewable materials, such as agriculture wastes, can be used as raw
materials for lactic acid production. Microorganisms belonging to bacteria and fungi are actively
involved in production of lactic acid from the provided raw materials. This article discusses
various raw materials, microorganisms, fermentation methods involved in production of lactic
acid, and also the applications of lactic acid in different fields.

1. Introduction capacity makes it useful as a moisturizer in

cosmetic formulations. Due to presence of
Lactic acid is of earnest importance as it is hydroxyl and carboxyl groups in its
not only used as a raw material for the structure, it can be converted into useful
production of various products by chemicals like esters, bio solvents, etc. (Gao
fermentation, but it is also used in food, C et al. 2011). Lactic acid is also used as a
pharmaceutical and textile industries monomer in the production of biodegradable
(Young-Jung Wee et al. 2006). It was PLA (Poly Lactic Acid) (Datta et al. 1995)
initially considered as a milk component by and it is used for medical purposes as in case
Scheele when he first discovered it in 1780. of prostheses, surgical sutures, etc. (Wee et
It was later named as Acide lactique by al. 2006). Due to its versatile applications,
Lavoisier in 1789 (H. Benninga 1990). CE lactic acid is deemed as one of the most
Avery was the first person to commercially important hydroxycarboxylic acids and it is
produce lactic acid in 1981 in Littleton, MA, in high demand (Lopes MS et al. 2012).
USA (Vickroy 1985). Around 370,000 metric tons of lactic acid
Besides its applications in food preservation, was produced in the year 2017 (C.
pharmaceutical and chemical industries, it is Rodrigues et al. 2017) and the demand of
also a widely used flavoring agent, acidulant lactic acid is rapidly increasing every year at
and inhibitor of bacteria. Its water retaining

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the rate of 5-8% (Yadav et al. 2011; 2. Raw Materials for lactic acid
Jamshidian M et al. 2010). production
Various fermentation approaches for the In order to produce large amount of lactic
production of lactic acid include batch, fed- acid by fermentation at low cost, cheap raw
batch, and continuous batch fermentations. materials are required. Besides being cheap,
Batch and fed-batch cultures yield high the raw materials should also have the
concentrations of lactic acid when compared properties such as ability to produce high
to continuous cultures. Though continuous yield, negligible or no formation of by-
cultures yield low lactic acid concentrations, product, high productivity, and less
the productivity is high (Hofvendahl et al. contamination so that not much pre-
2000). Microorganisms that are capable of treatment is required (John et al. 2009). The
production of lactic acid are of two groups: use of refined materials, such as
Bacteria and fungi (Litchfield 1996). carbohydrates, may reduce the product
Although fungal fermentation has a slight purification cost considerably, but it is not
advantage over bacterial fermentation as economical as it results in high production
filamentous fungi need only a simple costs (K. Hofvendahl et al. 2000). The
medium to produce lactic acid, by the usage following are some of the cheap raw
of glucose aerobically (Tay et al. 2002), it material sources for the production of lactic
requires vigorous aeration (Yin et al. 1997). acid: Starchy materials such as corn, potato,
Lactic acid can be also produced by rice, and wheat starch, and cellulosic
chemical synthesis. The chemical synthesis materials such as wood, cellulose are mainly
of lactic acid is mainly done by hydrolysis used as raw materials as they are in available
of lactonitrile by strong acids, which ample amount and cheap (K. Richter et al.
produces a racemic mixture of L- lactic acid 1998; K.V. Venkatesh 1997; C. Åkerberg
and D- lactic acid. Various other processes and G. Zacchi 2000). Starchy materials have
for production of lactic acid by chemical α (1,4) and α (1,6) linked glucose in the
synthesis include oxidation of propylene structure (Richter et al. 1994; K. Hofvendahl
glycol, nitric acid oxidation of propylene, et al. 1997). Cellulosic materials have β
etc. But none of the chemical synthesis (1,4) glucan, lignin, arabinan, galactan and
processes (except hydrolysis of lactonitrile) xylan (Litchfield et al 1996; Hofvendahl et
are economically and technically feasible. al. 2000).
The major advantage of fermentation
process over chemical synthesis is that it Apart from starchy and cellulosic materials,
requires cheap raw materials like starchy renewable sources such as agricultural
waste, molasses and other materials rich in residues, which are rich in carbohydrates are
carbohydrates (Anuradha et al. 1999). used. Agricultural residues are abundant as
about 3.5 billion tons of agricultural residues
The recent trend in usage of lactic acid is in are produced per annum, however,
the production of Poly Lactic Acid (PLA). availability is not the only criteria, price and
PLA could be potential replacement for purity also matter. The presence of
fossil fuel based plastics, but its production cellulosic residues in the agricultural
cost should be reduced to half of its current residues results in low protein content and
price in order to achieve that (Lopes MS et poor digestibility. So, the utilization of
al. 2012; Abdel-Rahman et al. 2013). If this agricultural residues is limited (John et al.
is made possible, the demand of lactic acid 2007). The following are some of the
would rise even higher. agricultural residues used for lactic acid

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production: corn cob (Vickroy 1985), 3 Microorganisms for lactic acid

cassava bagasse (Rojan et al. 2009), beet production
molasses (Ch. Kotzamanidis et al. 2002;
Yekta Göksungur, Ulgar Güvenç 1999), Microorganisms play a pivotal role in the
lignocellulose (Karel et al. 1997), production of lactic acid and they must be
cellulose(K.V. Venkatesh 1997), sugarcane readily available and cheap. Lactic acid
press mud (S. Xavier,B. K. Lonsane 1994), producing microorganisms are classified
and carrot waste(Ashok Pandey et al. 2001). into bacteria, fungi, and yeast. Most of the
It is reported that waste paper is a good lactic acid production is done industrially by
source as well (Yáñez et al. 2005; Park et al. the use of lactic acid producing bacteria (Cui
2004). et al., 2011; Kleerebezem and van
Loosdrecht, 2007; Nancib et al., 2009).
Whey and molasses, which are industrial However, the fungal species of Rhizopus
wastes, are commonly used as substrates for have their own advantage as they make use
lactic acid production. Whey contains of glucose aerobically to produce lactic acid.
lactose, protein, fat and mineral salts as it is But, the production rate of fungal
a by-product of dairy industry. fermentation is low due to mass transfer
Microorganisms such as Lactobacillus limitations (Park et al. 1998). The Genetic-
helveticus (A.W. Schepers et al. 2002; U. engineering techniques are exploited to
Kulozik and J. Wilde 1999) and improve the lactic acid yield and optical
Lactobacillus casei are used in production of purity by various microbial producers
lactic acid from whey (A.O. Büyükkilci and (Okano et al. 2010).
S. Harsa 2004; T. Pauli and J.J. Fitzpatrick
2002). Molasses, on the other hand, contains 3.1 Bacteria
large amount of sucrose as it is a waste
product from sugar manufacturing process. Lactic acid producing bacteria are classified
For the production of lactic acid from into four main categories, which are, Lactic
molasses, microorganisms like Lactobacillus Acid Bacteria (LAB), Escherichia coli,
delbrueckii and Enterococcus faecalis are Corynebacterium glutamicum, and Bacillus
used (Monteagudo et al. 1997; Göksungur strains (Litchfield et al. 2009; Budhavaram
and Güvenç 1999; Wee et al. 2006). For any and Fan 2009). Out of these, Lactic acid
fermentation process, it is essential to bacteria are most commonly exploited.
provide nutrients to the fermentation media. Choosing a proper strain is very important
Though it results in high production costs, because factors such as yield, productivity,
yeast extract is the most commonly used purity and nutrition requirements are
nutrient source for the production of lactic dependent on it (Rodrigues et al. 2017).
acid (Vickroy 1985; Hofvendahl et al.
Some of the limitations of lactic acid
2000). H. Oh et al. attempted to use corn
production by bacteria include low yield due
steep liquor as an alternative for yeast
to formation of by-product, requirement of
extract as it results in significantly low
nutrient rich medium, high risk of cell lysis,
production costs and succeeded. Malt sprout
necessity of mixed strains for development
extract, and whey permeates can also be
of phage-resistant strains to prevent
used as nitrogen sources in lactic acid
bacterio-phage infection (Rodrigues et al.
production (Suzanne F. Dagher et al. 2010).
2017).

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Lactic acid bacteria can produce lactic acid acid bacteria, some fungal species, such as
by anaerobic glycolysis with high yield and Rhizopus, with their amylolytic enzyme
productivity. They are present in dairy activity, can convert starch into L(+) lactic
products, meat, and in plants. Different acid (Wee et al. 2006). Some other
bacteria grow at different conditions. In advantages of fungal fermentation over
general, the optimal pH range for the growth bacterial fermentation include low-cost
of bacteria is 3.5–9.6 and the optimal downstream process, low nutrient
temperature is 5–45 °C (Rodrigues et al. requirements, and formation of fungal
2017). Based on the fermentation end biomass, which is an important by-product
product, Lactic acid bacteria are grouped (Zhang et al., 2007). Fungal fermentation
into two types: homofermentative and uses chemically defined medium, and so, the
heterofermentative. Homofermentative purification of products is simple. This is a
lactic acid bacteria glucose exclusively into major advantage in food industry (John et al.
lactic acid by Embden-Meyerhof pathway 2007). Generally, ethanol and fumaric acid
(Yun et al. 2003). Hence, homo fermentative are the common by-products formed by
LAB are used in commercial production of fungal fermentation (Litchfield, 2009; Vink
lactic acid. Some of the homo fermentative et al. 2010). The organisms of genus
lactic acid bacteria used in the production of Rhizopus are deemed as the best fungal
lactic acid are Lactobacillus delbrueckii, source for lactic acid production by fungal
Lactococcus lactis, Lactobacillus casei, fermentation (Rojan et al. 2007). Besides
Lactobacillus helveticus, and Lactobacillus Rhizopus, other fungi, like, the organisms of
acidophilus (Rojan et al. 2007). genus Monilia and Mucor are also used in
Heterofermentative LAB produce less yield lactic acid production (Prescott and Dunn,
due to formation of by-products. 1959). The main drawback of lactic acid
Lactobacillus pentosus (Bustos et al. 2004), production by fungi is that the lactic yield is
Lactobacillus bifermentans, and reduced as the carbon is utilized for the
Lactobacillus brevis (Cui et al. 2011) are production of by-products besides lactic
some of the examples of heterofermentative acid. The limitations of production of lactic
bacteria. Enterococcus mundtii (Abdel- acid by fungi also include mass transfer
Rahman et al., 2011) and genetically- limitation -which results in low production
engineered Lactobacillus plantarum (Okano rate, and requirement of vigorous aeration as
et al., 2009) have the capability to convert it is an aerobic process (Wee et al. 2006;
pentose sugars into lactic acid by Park et al. 1998).
homofermentative process. One of the key
reasons for usage of lactic acid bacteria in 3.3 Yeasts
industries is because it does not have any
All the fermentation processes require
adverse health effects (Rahman et al. 2013).
abundant amount of nutrient supply. In
The properties such as high acid tolerance
many of the fermentation processes, not
and the ability to be engineered for selective
only lactic acid, yeast is used as the key
production of D-or L-lactic acid make lactic
nutrient source. The major advantages of
acid bacteria commercially useful
using yeast as nutrient source include their
(Rodrigues et al. 2017). tolerance against low pH (1.5), which
3.2 Fungi prevents the regeneration of precipitated
calcium lactate, thereby reducing the cost of
Though majority of the lactic acid neutralization by neutralizing agents such as
production activities are performed by lactic calcium carbonate, and their ability to grow

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in mineral media (Rahman et al. 2013). The or product inhibition, and as the amount of
yield of lactic acid produced by fermentation nutrients provided is limited, low cell
with wild-type yeast as nutrient source is concentrations are obtained (Kadam et al.
low. With the advent of genetic engineering, 2006; Yun et al. 2003). Batch fermentation
genetically modified yeasts capable of is mainly of two types, which are, Solid
producing high yield of lactic acid have State Fermentation (SSF) and Separate
been created and they are being used Hydrolysis and Fermentation (SHF).
successfully (Bianchi MM et al. 2001). The
yeast of species Saccharomyces, Candida, Solid State Fermentation (SSF) is a process
Zygosaccharomyces, and Pichia are that occurs with no water or negligible
genetically modified to produce high yield amount of water. Natural raw materials such
of lactic acid (Rahman et al. 2013). The as wheat bran, rice bran, barley, fruit pulps,
main drawback of using yeast as a nutrient sugarcane bagasse are used as carbon source
source is that it leads to increase in in this process (Pandey and Ashok 2008).
production costs. However, corn steep liquor This process is used for the production of
(H. Oh et al. 2005), rice bran, and wheat pharmaceutical products, industrial
bran can be used as alternatives for yeast chemicals, feed, and fuel. Soccol et al.
(Yun et al. 2004). (1994) were able to produce 137.0 g/l of
lactic acid at the rate of 1.38 g/l/h by using
4. Fermentation methods for lactic acid Rhizopus oryzae by SSF. The usage of in
production solid state fermentation process produced
the lactic acid yield of 0.97 g/g of recycled
Fermentation of lactic acid, like any other paper (Marques et al. 2008). The advantages
fermentation process, is dependent on of using solid state fermentation method
factors such as raw materials used, nutrients include high productivity, single reaction
present in media, and the microorganisms vessel, rapid processing time, and less
used. Three different methods of enzyme loading (Abdel-Rahman et al.,
fermentation are practiced, namely, Batch 2011). In separate hydrolysis and
fermentation, Fed-batch fermentation, and fermentation process, the raw materials are
Continuous fermentation. first pre-treated and the unnecessary
compounds, such as lignin in the case of
4.1 Batch fermentation
lignocellulosic biomass, are eliminated.
In batch fermentation, all the required Then, the raw materials are subjected to
materials such as carbon source, nitrogen enzymatic saccharification and the
source and other components are added prior hydrolysate formed is subjected to
to beginning of the fermentation process. It fermentation (J. Choudhary et al. 2016). As
is the most commonly practiced SHF is preceded by such a hefty process for
fermentation process as it is simple to pre-treatment of raw materials, the real
perform. The major advantage of batch productivity decreases (Rahman et al. 2013).
fermentation is that it prevents It was reported by Marques et al. (2008) that
contamination to a good extent when the same Lactobacillus rhamnosus, which
compared to the other methods as it is a produced lactic acid yield of 0.97 g/g of
closed system, and so, high concentrations recycled paper by SSF produced only 0.81
of lactic acid is produced (Hofvendahl and g/g when done by SHF method.
Hägerdal, 2000; Rahman et al. 2013). The
drawbacks of batch fermentation include
low productivity due to substrate inhibition
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4.2 Fed-batch fermentation reported the production of lactic acid at the

rate of 1.56 g/l/h using Enterococcus
In Fed-batch fermentation, all the required faecium by continuous fermentation (Shibata
raw materials such as carbon source, et al. 2007). Ahring et al. reported the
nitrogen source, and other required production of lactic acid using Bacillus
components are added during fermentation coagulans (strain AD) at the productivity of
process at regular intervals of time without 3.69 g/l/h using continuous fermentation
removal of fermentation broth (Ding and (Ahring et al. 2016).
Tan, 2006). This type of fermentation is
especially useful to maintain low substrate 5. Applications
concentration by supplying nutrients to the
fermentation culture which in turn reduces Lactic acid is a hydroxycarboxylic acid with
substrate inhibition (Rahman et al. 2011). wide range of commercial and household
Ding and Tan reported that lactic acid applications. In the food industry, it is used
production by Lactobacillus casei using fed- as a preservative, flavoring agent, pH
batch fermentation was found to be more regulator, to increase shelf life, and to
efficient in production of lactic acid than control pathogens, among other applications.
other methods. They used 1% yeast extract As a food preservative, it inhibits putrefying
and glucose as raw materials and obtained bacteria, which prevents spoilage of food.
maximum lactic acid concentration of 210 Lactic acid, in the form of sodium or
g/l and L-lactic acid concentration of 180 g/l potassium lactate, is used to extend the shelf
at the rate of production 2.14 g/l/h, and the life of meat, poultry & fish. Due to its mild
yield of about 90.3% of L-lactic acid was acidic taste, lactic acid is used as an
obtained. Dong-Mei Bai et al. (2003) used acidulant in pickled vegetables, baked
Lactobacillus lactis for the production of L- goods, salads, and beverages. In case of
lactic acid and obtained about 97% yield of dairy industry, lactic acid is an excellent
L-lactic acid at the rate of 2.2 g/l/h. acidification agent due to its natural
presence in dairy products, combined with
4.3 Continuous fermentation the dairy flavour and good antimicrobial
action of lactic acid. It is also used to
Continuous fermentation involves addition enhance savoury flavours. Lactic acid is
of fresh medium to the fermenter while used in chocolates and sweets for the
withdrawing the already existing broth at the flavour, as well as for obtaining the correct
same rate. Due to this, the concentrations of pH. Other advantages of adding lactic acid
substrates and products is maintained in chocolate and candy production include
constant (Rahman et al. 2013). The ease of handling, low inversion rate, and
advantages of continuous fermentation production of clear candies (Wee et al. 2006;
include prevention of end-product Corbion Purac).
inhibition, less frequency to shut down, less
decrease in productivity during lag phase, In pharmaceutical industry, lactic acid is
inoculation of culture is done once only, used as an electrolyte in various
high product yield can be obtained, saves parenteral/intravenous solutions. These
time and involves less labour work. parenteral solutions are prepared to
Continuous fermentation suffers from a few supplement bodily fluids. In addition to this,
drawbacks such as contamination, lactic acid comes into play in pH-regulation,
requirement of field operator with expertise, chiral intermediate and metal sequestration
and it is expensive to perform. Shibata et al. (Ramzi A et al. 2015). Lactic acid is also

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 Battula Savithra Krishna et al. / International Journal of Biotech Research (2018)

involved in mineral preparations, tablets, hygiene products (Martinez et al. 2013). The
prostheses, controlled drug delivery system, recent advent in lactic acid application is
surgical sutures, and in preparation of Poly Lactic Acid (PLA). It is a
dialysis solutions for dialysis processes like biodegradable plastic and has numerous
Continuous Ambulatory Peritoneal Dialysis applications in day-to-day activities such as
using artificial kidney machines (Wee et al. food packaging, containers, trash bags,
2006). Lactic acid, along with its salts, acts protective clothing, etc. (Södergård and Stolt
as an intermediate in the manufacture of 2002; Vink et al. 2003).
pharmaceuticals, to adjust the pH of
preparations. Pharmaceuticals contain L (+) 6. Conclusion:
lactic acid as the D (-) isomer is not
Lactic acid is widely used in
metabolized by the human body. Salts of
pharmaceutical, food, chemical and
lactic acid such as Calcium, iron, sodium,
cosmetic industries due to its easy
and other salts are used in pharmaceutical
availability of raw materials, high
industry because of their anti-tumour
productivity, and low cost of production. It
activity (Ramzi A et al. 2015).
is commercially produced in batch and
In the chemical industry, lactic acid is used continuous methods. However, batch
as neutralizer, cleaning agent, descaling fermentation method gives high
agent, pH regulator, and antimicrobial agent. concentration, continuous fermentation
Lactic acid is an excellent remover of gives more productivity. In batch
polymer and resins due to its high solvency fermentation lactic acid is produced by
power (Wee et al. 2006). Due to presence of bacterial genus Lactobacillus and fungal
two reactive functional groups, that is, a genus Rhizopus. Genetically produced yeast
carboxylic group and a hydroxyl group, species Saccharomyces, Candida,
lactic acid can undergo a variety of chemical Zygosaccharomyces, and Pichia are found to
reactions that yield useful chemicals. The produce high yield of lactic acid but cost of
following are the chemicals that lactic acid production is high. Batch fermentation is
gets converted into when it undergoes carried out by Solid State Fermentation
various reactions: (SSF) is found to be more effective than
Separate Hydrolysis and Fermentation
Lactic acid has numerous applications in the (SHF). Refined starch and cellulose
field of skin care and cosmetics. Studies materials are commonly used as raw
have found that lactic acid plays an essential materials for lactic acid production, however
role as a skin brightener and it also aids in the present trend of research is towards the
removal of brown spots on skin (M Ochi et use of renewable resources such as
al. 2004). Lactic acid acts as a moisturizer, agricultural waste materials: corn cob, corn
anti-acne agent, humectants, anti-tartar stalks, bagasse, beet molasses etc. In this
agent, pH regulator, skin-lightening agent, study it is revealed that Fed-batch
and skin-rejuvenating agent. Lactic acid acts fermentation using lactobacillus produces
as a moisturizer due to its water retaining high yield of lactic acid compared to other
capacity. Lactic acid promotes collagen methods. There is a lot of scope for the
production, and hence it prevents wrinkles production of lactic acid using renewable
and fine lines on skin and keeps the skin materials as raw material sources.
firm. So, it acts as an anti-aging tool
(Bouwstra and Ponec 2006). Lactic acid also
has applications in manufacture of oral

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Lactic Acid
Raw Material Microorganism Reference
Yield

Corn Enterococcus faecalis RKY1 63.5 g/L H. Oh et al 2005

Lactococcus lactis ssp. lactis ATCC 19435 106.0 g/L K. Hofvendahl et al. 1997
Wheat
Enterococcus faecalis RKY1 102.0 g/L H. Oh et al. 2005
Molasses Lactobacillus delbrueckii NCIMB 8130 90.0 g/L C. Kotzanmanidis et al. 2002
Lactobacillus coryniformis ssp. torquens
Cellulose 24.0 g/L R. Yáñez et al. 2003
ATCC 25600
Lactobacillus helveticus R211 66.0 g/L A.W. Schepers et al. 2002
Whey
Lactobacillus casei NRRL B-441 46.0 g/L A.O. Büyükkilci et al. 2004

Rice Lactobacillus sp. RKY2 129.0 g/L J.S. Yun et al. 2004

Wood Lactobacillus delbrueckii NRRL B-445 108.0 g/L A.B. Moldes et al. 2001

Apple pomace Lactobacillus rhamnosus ATCC 9595 32.5 g/L Gullón et al. 2008

Glycerol E. coli AC- 521 56.8 g/L Hong et al. 2009

Lignocellulose- 129 g/L Abdel-Rahman MA et al.
Enterococcus mundtii QU 25
derived sugars 2015
Raw sweet Lactobacillus paracasei and Lactobacillus
198.32 g/L Nguyen et al. 2013
potato coryniformis
Table 1: Raw materials used in lactic acid production by fermentation and their yield

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Field Applications

 Preservatives
 Acidulants
 pH regulators
Food industry
 Mineral fortification
 Bacterial inhibition
 Dialysis solution
 Mineral preparations
 Prostheses
Pharmaceutical industry  Surgical sutures
 Controlled drug delivery systems
 Parenteral/intravenous solution
 Neutralizers
 Chiral intermediates
 Green solvents
Chemical industry  Cleaning agents
 Descaling agents
 pH regulators
 Moisturizers
 Anti-acne agents
 Humectants
Cosmetic industry  Anti-tartar agents
 pH regulators
 Skin-lightening agents
 Food containers
 Protective clothing
Poly Lactic Acid
 Trash bags
 Rigid containers
Table 2: Applications of lactic acid in various fields (modified after Wee et al. 2006; Vink et al. 2003)

Reaction Chemical produced

Hydrogenation Propylene oxide

Decarboxylation Acetaldehyde

Dehydration Acrylic acid

Reduction Propanoic acid

Condensation 2,3-pentanedione

Self-esterification Dilactide
Table 3: Chemicals that lactic acid gets converted into via various reactions (Varadarajan et al. 1999)

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