B

IOD IVER S ITY EVALUATION-B OTANI CAL PERS PECTIVE

17

-28, 2012

Edited by N.S. Atri, R.C. Gupta, M.l.S" Saggoo E V.K. Singhal Published by Ws Bishen Singh Mahenilra Pal Singh, Dehra Dun, (India)

MICROALGAE AS A POTEI\TIAL SOURCE FOR BIODIESEL T'RODUCTION

|itender, {,J"8" Singh and A.S. Ahluwalia* Department of Botany, Panjab Llniaersityt Chandigarh-1"6A AM
*Email : phyko s@pu. ac.in Self sufficiency in energy requirements is critical to the success of any growing economy. With the ever increasing demand for diesel and environmental squalor caused by burning of diesel fuel, research for the development of a sustainable alternative fuel is being carried all over the world. Sustainable production of renewable energy is being hotly debated globally since it is increasingly understood that first generation biofuels, primarily produced from food crops and mostly oil seeds are timited in their ability to achieve targets for biofuel production, climate change mitigation and economic growth. These concerns have increased the interest in developing second generation biofuels produced from non-food feedstock such as microalgae, which potentially offer greatest opportunities in the longer terrn. Biodiesel has become an attractive alternative to diesel fuel in last many years. Microalgae are considered a very good feedstock for biodiesel production due to their very high yield and their no competition with food crops. Oil productivi$ of many micro-algae greatly exceeds the oil productivif of the best oil producitg crops. Major steps in harvesting this potential include inputs from research u.d technologies along with monet ary inputs. Investing in R&D for biomass energy is investing in our farmers, environment, economy that is, in our future"

K"y Word s : Mic r o algae,B

io d ies el,

Transesterific ation.

INTRODUCTION
The world is entering into a period of declining non-renewable energy resources; populariy know as "Peak Oil", while energ"y demand is increasing (Campbell, 2008). Continued use of petroleum based fuels is now'widely recogni zedas unsustainable because of depleting supplies and contribution of these fuels to pollute the environment. Over reliance on petroleum derived fuels has caused carbon dioxide (COr) enrichment of the atmosphere which

has already exceeded 450 ppm Cq* (e = equivalent contribution of all greenhouse gases) , as already being in the dangerously high range and this threshold was exceeded 10 years earlier than had previously been predicted (IPCC,2007). The annual mean growth rate far 2007 was2.14 ppm - the fourth year in the past six to see an annual rise greater than 2 pp* (The Guardian 12th May, 2008).

18

Iv[icroalgae

GS

a Potential Biomass For Biodiesel."

It is expected that with the development of new growing economies, such as India and China, the global consumption of energy will raise and lead to more environmental damage (IEA, 2007). India, like any other developing country, relies on energy for sustained growth and development and consequenfly depends on crude petroleum imported from other countries to cater to the large demands of fuel in the automobile, industrial and agricultural sectors. India produced 34.11 million tones of crude oil in 2007 - 08, and imp orted l2'1,.67 million tones of crude oil worth Rs.2726.99 brlhon. In India, emissions from energy use in transport and industrial sectors are higher than in agriculture sector (Naravana et al., 2009). The researchers have the daunting challenge to find out sufficient supplies of clean energy for the future, which is intimately linked to global sustainability, economic prosperity, and quality of life (Campbell,2008).
Indian economy is essentially diesel driven and the consumption of diesel fuel is four to five times more than motor-gasoline which is characteristically differentfrom several developed economies. Therefore, in Indian context, there is an urgent need to identify and commercialize renewable alternative fuels for diesel substitution. Biodiesel has become a sustainable substitute to diesel fuel as biodiesel is produced from vegetable oil or animal fat through a chemical process known as transesterification. Research on differentaspects of biodiesel production and utilizationin diesel engine has been reported by a number of researchers (Basha et a1.,2009; Jitender et a\.,2009;Li et a\.,2009; Nabi et a1.,2009; Oner and Alturu 2009; Patil and Deng, 2009; Sharma and Singh,2009;Yan et aI.,2009). Biodiesel can be used u" Litf,"t direct substitute or as an extender to fossil diesel fu'el in compression ignition engines and has the potential to reduce carbon dioxide, hydrocarbon and carbon monoxide emissions. Biodiesel is currently produced from the oil synthesized by conventional fuel crops like latropha and Karanja that harvest sun's energy and store it as a chemical energy. In Indian contex! latrophais the most potential crop for biodiesel production. latropha curcas is a hardy plant and can even thrive on waste and degraded land (Kumar and Sharma,2005).

BIODIESEL AND ITS ADVANTAGES Biodiesel is a mixture of fatty acid alkyl esters obtained by
transesterification (ester exchange reaction) of vegetable oils or animal fats. These lipid feedstocks are composed by 90 - 98% (weight) of triglycerides and small amounb of mono and diglycerides, free fatty acids (1 - 5%), and residual amounts of phospholipids, phosphatides, carotenes, tocopherols, sulphur compounds and haces of water (Bozbas,2008). Transesterification is a multiple step reaction, including three reversible steps in series, where triglycerides are converted to diglycerides, then diglycerides are converted to monoglycerides, andmonog$cerides arethenconvertedto esters (biodiesel) and glycerol (by-product). The overall transesterification reaction is described

'fitender,

U"B. Singh andA.S. Ahluwalia

R1-,

19

in Figure l-, where the radicals known as fatty acids"

R2, R3 represent long chain hydrocarbons,

:ll CH:-O-C-Rl
o
CII
catsll'st

(!

o
Rt

* Co

6Rt

Cllr

flfi

-o-e-Rr t
ll

3R'oH

F*. lh-c-oR' +
0

CII

*. 0ll

Cru'-0-C-Rs
Triglyccridcs Alcohol

il

R3-C-ORt
Estcrs

cHr

0H

GI"yccrol

Fig.l: Transesterification of triglycerides (overall reaction) Primary advantages of biodiesel are that it is one of the most renewable fuels and also non-toxic and biodegradable (Gerperg 2005). Today, biodiesel has come to mean a very specific chemical modification of natural oils. The use of vegetable oils as altemative fuels has been around for 100 years when the inventor of the diesel engine Rudolph Diesel fust tested peanut oil in his compression ignition engine (Shay, 1993). Oilseed crops such as rapeseed and ioybean oil have been extensively evaluated as sources of biodiesel' One of the biggest advantages of biodiesel compared to many other alternative transportation fuels is that it can be used in existing diesel engines without

modification, and can be blended in at any ratio with peholeum diesel. Biodiesel produced fromvarious vegetable ofu haveviscosities close to those of diesel. The volumehic heating vilues are little lower, but they have high flash points. The performance of diesel engine with biodiesel is generally comparable to that of conventional diesel fuel with reduced engine emission
of particrrlates, hydrocarbons and carbon monoxides. Since the characteristics of biodiesel are similar to those of diesel, the former is a strong candidate to

replace diesel in the future.

Inview of the environmental considerations, biodiesel is considered as 'carbon neutral' because all the carbon dioxide (COr) released during consumption had been sequestered from the atmosphere for the growth of vegetable oil crops (Barnwal and sharma, 2005). Commercial experience *ith biodi"sel his been very promising. Biodiesel performs as well as petroleum diesel, while reducing emissions of particulale m.after, carbon monoxide (Co), hydrocarbons and oxides of sulphur (sox) (Barnwal and Sharma, ZObS). Emissions of oxides of nitrogen (NOx) are, however, higher for biodiescl in many engines. Biodiesel virtually eliminates the notorious

20

Ivlicroalgae as n Potential Biamass For Biodiesel..,

black soot emissions associated with diesel engines and the total particulate

matter emissions are also much lower. Other environmental benefits of biodiesel include the fact that it is highly biodegradable and appear to reduce
emissions of air tsxics and carcinogens (relative to petroleum diesel). usage of biodiesel will allow a balance to be sought between agriculture, economic development and the environment.

The'centerpiece of India's plans for biodiesel development and com.mercialization was the National Biodiesel Mission (NBM), formulated by the Plannirig Commission of Government of India. Based on extensive reseafch carried out in agricultural research centers, it was decided to use
latropha carcas oilseed
aS

the major feedstock for India's biodiesel programme.

NBM was planned for two phases. Phase I was termed as demonstration phase and has been carried out from year 2003 to 2007 (Planning bomrnission, 2003). The woik done during this phase were development of land, latro,phaoilseed nurseries, cultivation of latropha on 400,000 ha waste and the centers, expression oil and, collection setting up of seed latropha

irrstallation of 80,000 Mt/year transesterification to produce biodiesel from latropka oil. Phase II planned with a selfsustaining expansion of the programme leading to the production of biodiesel to meet 20 % of the county's diesel requiremenb by 201L-2u12. The lack of assured supplies of vegetable oil feedstock has foiled efforts by the private sector to set up biodiesel plants in India. Commercial biodiesel production has not yet started in India. So far cnly two'firrns, Naturol Bioenergy Limited (NBL) and Southetn Online BiotiEe&nslogies, have'enbarked or,t biodiwel projects, both in the southern statedf AndhraFradesh. Naturol'Bitlenirgyliorited'(NBl), a jointventure with the Austrian biodiesel firm Energea Grnbh and the investment firm Fe Clean Energy (USA), has planed to install a 300 tonnes /day (t/d)/(90,000 tonnes/year) (t/fl biodiesel plant in Kakinada, Andhra Pradesh' So, there are many conshaints for the biodiesel production in India and phase I of NBM has not given the anticipated resulb. Recent research and development

on microalgae has exposed a new road ahead that can be promising

sustainable feedstock for biodiesel production,

MICROATGAE, AN EMERGING FEEDSTOCK FOR BIODIESEL

PRODUCTION
Biodiesel from microalgae seems to fe the only renewable biofuel that has the potential to completely displace petroleum-derived transport fuels withouiadversely affecting supply of food and other crop products (chisti, 2008). Algal ponds and bioreactors for algae production are situated on nonarable land, h owever,latropfta is mainly grown on marginal land. Availability of feedstock for biodiesel production has always been a matter of concern and challenge for biodiesel industry and in this context, yield of biodiesel crop should be as high as possible for biodiesel industry to survive. Table-1 summarizes the yield per acre of various feedstocks.

litender, U.B. Singh and A.S. Ahluutalia

21

Table-l: Comparison of microalgae with other biodiesel feedstocks.

Plant source
Seed

oil

Oil yietrd Land use

Biodiesel

content (% oil by :wt" in biomass)

(L

oIL/

(r*

year

ha year)

kg biodiesel)
66 31 18 15

productivity
(kg biodiesel/ ha
year)

CornlMaize (Zea mays L.) Hemp (Cannabis satiua L.)

Soybean (Glycine max L.) Jatropha {lntroplta ntrcas L.) Camelina (Cnrnelfuta satiua L.)
C-anola / Rapesee

44 33 18 28

1,72

152
321

363 636 741

91,5

562
656
809

42
L.) 41

72
12
1.1

d (Brassiu nayts

974 1070
1,307

862
946 1L56
47 47

Sunflow er (Heliantlrus anrunts L.)

40 48 36 30 50 70

Castor (Ricinus comtnunis)

Palm

otl (Elaeis gttineensis)

5366
58,700

Microalgae (low oil content) Microalgae (medium oil content) Microalgae (high oil content)

0.2
0.1 0.1

5t,927 I6,515
121,1.94

97,800 136,900

(Source: Mata et al., 2070)

Microalgaehre bunlight- driven cell factories thatconvert carbon dioxide to potential biofuels, foods, feeds and high value bioactives (Chigti,20OZ1,: They cornprise a l,arge and diverse group of photosynthetic organisms that exhibit a variety of habitats in temperate, tropical and polar regions. Most commonly, these organismsflourish in aquatic habitats. However, they can also grow in soil, deserb, oil-fields, bare rocks, hot springs, etc. (Dhingra and

Ahluwalia, 2007).It contains lipids and fatty acids as mernbrane components, storage products, metabolites and sources of energy. Algal strains, diatoms and cyanobacteria (categorized collectively as "Microalgae") have been found to contain proportionally high levels of lipids
(over 30%). Microalgal biomass, like other plant biomass, is potentially suitable for conversionto liquid (gasoline, biodiesel, ethanol) and gaseous
(methane and hydrogen) fuels. Most of the oleaginous microorganisms like

microalgae, baciilui, fungi and yeast are all available for biodiesel production (Meng et al., 2009).

Microalgae have a number of unique benefits. As aquatic species, they do not require arable land for cultivation. This means that algal cultivation does not need to compete with agricultural commodities for growing space. In fact, algal cuitivjation facilities can be built on marginal land that has few other useJ. Algae also have a greater capacity to absorb CO, than land plane, lnd are not pione to photosynthetic inhibition under conditions of intense ;unlight. Afier oil extraction from algae, the remaining biomass fraction can re nr6d as a high protein feed for livestock which gives further value to the

22

lvficron\gne frs n Fatemtin| Biomnss For Bi*diesel.""

2008)' Microalgae apPear process and red.uees waste to even zero (Campbell,

tobetheadvantageofcostsdependingo:ritsqp6rabilityofhigher

rate compared photosynthetic efficiency, larger biomass and faster growth Si % of its dry is usually [o tf,ut f f ol crops. Oil conteni of many microalgae

*"ight 1so"g et il.,ZOO}l.Oepending on species, microalgae produce oils are tirrfi, .i fipias, hydrocarbons ana otnei complex,oils. Not all algal usingconunonly. occur oils ,"ituuru r* *aking biodiesel, however, suitable of producfion the with *i.ioufguu to proiuce biodiesel will not interfere strains micro-algal The food, foider uod oth", ptoducts derived from crops. and are of great *i,iit i*h oil or lipid content are summarized in Table-2 of biodiesel production the for i.rt r"riio tt u q.roi fo, u r*tainable feedstock
(Chisti,2A0n.
Table*?: Oil e#ffitemt of $offite microalgae"
fuf ie
S S

differerrt

roalgae
*bliEutws deswlws dimorPkus
wwlgffiYds

mig c*ffi#emt {% dry weighft}

eewe desffiE &s

cene

Cklsrelln C\il*rella

BotryoeCIrcer.ds s$3.

$p"

{ryp

tke

ert

diwitr W gyAW:

Cyiliwdrst'iqeew_f,ff:

ffiMwnldelfn priwr.l#{te
fsmelaryses
tvfl.sw s$3.

rullgnwtksa

sre

Js

ru m

FJmrereoelafores

sP.

(Source; Chisti,2007)
as a source There are many advantages of microalgae over h$her plants

of kansportation biofu els like:

r r

neuhal lipids/ Microalgae synthesize and accumulate large quantities of rates (e.g. 1-3 high oil t20-50% dry celt weight (Dcw)l and grow at

doublings/daY).

oil yield

per unit area of microalgal cultures could greatly exceed the

yield of best oilseed croPs.

oMicroalgaecanbecultivatedinsaline/brackishwater/coastalseawater with conventional on non-arable land, and do not compete for resources
agriculture.

semiarid lands) Microalgae tolerate marginal lands (e.g" desert, arid and that areirot suitable for conventional agriculture'

fitender, 1/"ts" 9irugl.' nndA.S" Ahlwwnhiq

23

Microalgae utilize nitrogen and phosphorus from a variety of

feed operations and industrial and municipal wastewaters), providing the additional benefit of wastewater bioremddiation.

wastewater sources (e.g. aglicultural run-off, concentrated animal

I r I

Microalgae sequester co, from flue gases emitted from fossil fuel-fired power plants and other sources, thereby reducing emissions of a major greenhouse gas. 1 kg of algal biomass requires about 1.8 kg of Ce.

Microalgae produce value-added co-products or by-products (e.g., biopolymers, proteins, polysaccharides, pigments, animal feed and fertilizer) and does not nebd herbicide and pesticide.

Microalgae grow in suitable culture vessels (photobioreactors) throughout the year with higher annual biomass productivid on an
area basis.

Microalgae are currently cultivated commercially for human nutriuonai, products around the world in several small- to medium-scale produ-ction systems, producing a smaller amount to several hundred tons of biomass annually. About half of this production takes place in mainland China, with the rest in Japan, Taiwan, U.S.A., Aushalia and India, and a few small
producers in some other countries as well (Benemann, 1997;2009).Microalgal

cultivation using sunlight energy can be carried out in open or covered ponds or closed photobioreactors, based on tubular, flat plate or other designs. closed systems are much more expensive than open ponds and present significant operating challenges (overheating, fouling, etc.) and due to gao
exchange limitations, among others, cannot be scaled-up much beyond about

a hundred square meters for an individual growth unit.

unlike the open ponds, photo bioreactors essentially permit single-species culture of
microalgae for prolonged durations. Photo bioreactors have been successfully used for producinglarge quantities of microalgal biomass (Chisti, 200n.Xi et al. (2006) extracted microalgal oil from the heterotrophic cells using n-

hexane and then hansmuted it into biodiesel by acidic transesterification" The heating value of biodiesel was found to be 41MJkg/L, the density as 0.86akg/L and the viscosity as 5.2x10a Pas (at 40"C) (Xu ef aL,20A6).

MICROATGAE BIODIESET VATUE CHAIN STAGES Although in a simplistic view, microalgae may not seem to differ
significantly from other biodiesel feedstocks, they are microorganisms riving essentially in liquid environments, and thus with particular cultivation, harvesting, and processing techniques that ought to be considered in order to efficiently produce biodiesel. All existing processes for biodiesel production from microalgae include a production unit where cells are grown/ followed by the separation of the cells from the growing media and subsequentlipids extraction. Ther; biodiesel or other biofuels are produced in a form akin to
existing processes and technologies used for other biofuel feedstocks. t{,ecently

24

Microalgae as a Potential Biomass For Biodiesel.."

species depending on local specific conditions and the design and implementation of cultivation system for microalgal growth. It is followed by the biomass harvesting, processing and oil exhaction to supply the biodiesel production unit.

other possibilities for biofuel production are being pursued instead of the transesterification reaction, such as the thermaf &acking (or pyrolysis) involving the thermal decomposition or cleavage of the triglycerides and other orghnic compounds presented in the feedsiock, in simpter molecules, namely alkanes, alkenes, aromatics, carboxylic acids, among others (Babu, 2008; Boatenget a1.,2008). Figure 2 shows a schematic representation of the algal biodiesel value chain stages, starting with the selection of microalgal

ECONOMIC FEASIBILITY OF BIODIESEL PRODUCTION

There is a common myth about biodiesel that it costs too much, but it is nothue altogether. The economic feasibility of biodieselproduction depends upon the fact that the feedstock biomass should be producible with lowinput (ie. less fert'lizer, pesticides and energy) on land with low agricultural value and requires low-input energy to convert feedstocks into biodiesel (Haas et al., 2006; Hill ef al., 2006). They also reported a direct linear relationship between the cost of feedstock and biodiesel production cost.

Algae and Site Selection

CO,
Nutrients

Algae cultivation

Algal culturc
(0,t)2{.1.06oh

nt TSS: total suspendetl solids)

Algal efliment (2-7% TSS)

Harvesting
Algal Slurry (5-15% TSS)

Biomass processing
(dewatering, thickenhg, filtering, dryng)
Nutrients recycle
Algal Cake (15-25% TSS)

il extraction
cell disruption and oil extraction)
Lipids and free fally acids

Biodiesel Production
Fig.2: Microalgal biodiesel value chain stages

litender, U.B. Singh andA.S. Ahluwalia

25

Ramesh et aI. (2007) calculated a net cost of production of latrophabiodiesel as Rs. Zi9 /1. There is, therefore, utmostneed to explore microalgae as alternate feedstock for biodiesel with minimal cost

It can be safely concluded that use of microalgae as a source of biomass for biodiesel production will be much cheaper than other plant crops, because many unicellular algae multiply their biomass within a short time. Thus use of microalgae for biodiesel production poses no problem atleast at economic level. Acclim at:rzrng microalgae for using flue gas in place of CO, for their growth can improve the economic feasibility of microalgal biodiesel through earning of carbon credits. The biggest and most important hurdle and utthzabion of microalgal fuel is the high cost and energy requirement" The forthcoming research and technology should concentrate on reducing the expenditure and improving the cost-effectiveness of microalgal fuel. These include strategies like selection of cost effective technologies for biomass harvesting and dryirg, high- value co-product skategy, best strain selectiory

metabolic engineering and systems biology approaches and linking microalgal production with wastewater treatment. Biotechnology should be used for maki^g biodiesel production from algae, a cheaper process. If s a time to go for highvalue lowvolume products frombiomass. Algalbiorefinery has a great potential to achieve this goal.

CONCLUSION
It can be said that algae are not just aquatic weeds rather these can prove to be the most potent raw material source for biodiesel production that too without endangering global food security and warming. Moreover, use of algae in biodiesel production can also act as a potential tool for carbon

sequestration and curbing climate change. But before these optimistic

assumptions come hue, there are certain challenges in fulfilling these dreams. These include extensive research trials / experiments for standardizing the protocol for specific algal species for ensuring continuous supply of biofuel, identification of environmental factors/stresses that can enhance oil/lipid accumulation in microalgae without affecting their growth rate, involvement of genetic engineering technology to increase the activity of enzymes related to lipid production and storage. Focused efforts in developing microalgae

with high lipid content for biodiesel production would be a new and
promising approach in biofuel production. There is utnnost need to develop ways for producing enormous amount of biomass for algal oils. Closed bioreactors save water in minimizing evaporafion whereas open race way ponds ufilize solar energy. Larger amount of microalgal biomass can also be achieved through various approaches involving improvement of algal strains, optimum lightquantity, quality and duratioru culture nutrienb, heterotrophy, temperature, etc. It is better to employ algal strains which normally grow in saline water because they accumulate more hydrocarbons and do not get contaminated even in the open ponds.

zfr

fuLir:ronlgae #s #

Fsterttial Bismsss

Fsy ffiisciesel",.

ACNOI,T/LEDGEMEI{TS
The authors are indebted to the university Grants Commission, New Delhi (A'S. Ahluwalia)forSAP-DRS-II grants and to rhe Council of Scientific and Industriar Research, New Derhi fliLnder and u.n. si"jr,jlr;;;ffi;;

financial assistance in the form of pi_Snn.

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A: General 360: 16g-170.