Agregando Valor aos
Coprodutos
Isabel Campos Portugal
Ben Gurion University of the Negev
Novembro /2015
http://web2.bgu.ac.il/algal/News.htm
O que são microalgas?
 Microalgas compõem um enorme grupo de microrganismos fotossintizantes
procariontes e eucariontes
 Podem habitar : ambientes de água doce, salobra e marinhos (solos/condições
extremas)
 Crescem rapidamente e podem suportar condições adversam devido a sua
estrutura unicelular/ multicelular
Spirulina platensis
Lobosfera Incisa
Chaetoceros teres
*Uma descrição bem aprofundada sobre microalgas ( Richmond ,2004)
Antecedentes históricos do
microalgas para fins comercias
1950
1960
1º- Cultivo em larga
escala Japão /
Tailandia
1970
USA- crise energética
1980
Chlorella ~ $11/kg -->
fermentação da biomassa
hundreds of tons (mainly
produção de metano
in Japan)
- Spirulina (Sosa
Aumento da população
Texaco) ~ 1000t/year
Busca por alternativas fontes
~10-15$/kg
proteicas
- Algae for
*Biomassa de Algas
aquaculture - Isochrysis,
Nanochloropsis
- Dunaliella for betacarotene
uso
de
1986
Dunalliela Salina
Coproduto? O que é coproduto?
 O mercado é virtualmente ilimitado:
 Números de espécie
 Estima-se 50.000 , mas so 30.000 sp
forma descritas / estudadas
 Tipo de cultivo
 Condicao de esstress (Nutri / luz/
salinidade) - lipídio, pigmentos
 Necessidade do Mercado
 proteina, pigmento, PUFA ou Betacarotena
Divisor de Águas - Importante publicação
 No artigo da FAO “ Algae-Based
Biofuels
Applications and Coproducts”
Produtos Bioenergéticos obtidos de
Microalgas
Esquema 1: Visão geral das opções de modo para produzir biodiesel a partir
de produtos de microalgas.
(FAO, 2010)
Opções não
energéticas à
base de algas
 Producao
de
microalga : 7000 tons/
dw (2004)
FAO(2010)
Mercado
Carotenoides
 Note-se que o preço
do produto à base de
algas é várias vezes
mais elevados
Tabela 03 : Mercado e preço carotenoide Dr Niels- Henrik Norker ( FAO, 2010)
O
mercado
dos
carotenoides
é
crescente, mas não
deverá
aumentar
dramaticamente
Gráfico 01 : Mercado Global de Carotenoides e futuro outlook em milhões
de dólares – BBC Researcch ( FAO, 2010)
Produtos de
Alto valor
 Table
06:
equilíbrio
econômico e ambiental
da produção de produtos
de alto valor por tipo de
algas
(
Carotenoide e Astaxantina
Cultivo de H. pluviavis
Commercial applications of microalgae
Chemical structures of microalgal pigments. (a) β-Carotene, (b)
astaxanthin, (c) phycoerythrin (in phycocyanin, the CH=CH2 group noted
an asterisk is replaced by CH3–CH2).
Journal of Bioscience and Bioengineering, Volume 101, Issue 2,
2006, 87–96
Algas como fontes de Farmacos
Indústria farmacêutica está crecendo a uma taxa de crescimento anual ( CAGR) de
cerca de 8%, enquanto o mercado farmacêutico mundial está previsto para chegar
a US $ 1043.4 bilhoes em 2012.
O mercado farmacêutico global expandida por um modesto de 3% em 2010.
Naquele ano, o total de vendas de produtos farmacêuticos em todo o mundo por
região geográfica foram os seguintes:
 EUA– 39%
 Europa – 29%
 Asia, Africa, and Australia – 15%
 Japão – 11%
 America Latina – 6%
Mercado Brasileiro
Produtos de alto valor agregados
Product Group
Phycobiliproteins carotenoids
Polyunsaturated Fatty acids
( PUFAs)
High - Value Products from Microalgae
Potential Application
Product
Phyconcyanim
Pigments , comestics , pro
Beta caratone
vitamin , pigments
Astaxanthin
Infant formulas for full-term
infants
γ- Linolenic acid GLA
Nutritional supplements
Microorganismo Producer
Spirulina platensis
Dunaliela salina
Hemantococus pluvialis
Spirulina
Infant formulas for full-term
infants /preterm infants
Nutritional supplements
Arachidonic acid AA
Parietochoris insisa
Porphyridium
Nutritional supplements
Eicosapentaenoic acid EPA
Chlorella minutissima
Nannochloropsis, Phaeodactylum,
Nitzschia,Pavlova
Aquaculture
Infant formulas for full-term
infants /preterm infants
Docosahexaenoic acid DHA
Crypthecodinium, Schizochtrium
Nutritional supplements
Aquaculture
Tabela 2: High value product from microalgae ( modified Li et al 2008 ; Spolare 2006)
Tabela 05: Ocorrencia de LC- Pufa em varias classes de microalgas
(Khozin-Goldberg et al., 2011)
Upscaling and optimizing DGLA
production by Lobosphaera incisa
∆5 desaturase mutant (P127) and
testing P127 as a dietary
supplement in fish
Fish Health Lab
Fundamentação teórica e Importância do
Projeto
Demanda do mercado para os ácidos graxos poliinsaturados de cadeia
longa ômega-3 e ômega-6. (LC-PUFA)
Algumas microalgas podem acumular grandes quantidades de LCPUFA sob a forma de triacilgliceróis (TAG).
L incisa e sua cepa mutante (P127), são produtores de grandes
quantidades de ômega-6 ARA e DGLA, respectivamente
DGLA afeta o metabolismo lipídico celular e
biossíntese eicosanóide.
P127 - DGLA de
uma fonte alga verde fotoautotrófico
Importância do DGLA / ARA
DGLA (20:3ω6)
ARA (20:4ω6)
DGLA: DGLA administration increased the ratio of PGE1/PGE2 in rat plasma =>
induces anti-inflammatory effect (Umeda-Sawada et al., 2006)
Anti-inflammatory effects were reported to include: anti-proliferative ; anticancer; (Wang et al., 2012); antithrombotic, antihypertensive, anti-allergic
and artherosclerotic activities (Kawashima et al.,2008; Takai et al., 2009)
ARA: A primary substrate for the biosynthesis of eicosanoids like prostaglandin
PGE2 and leukotriene B4 involved in inflammatory and immune responses
(Calder 2008; Lee et al., 2009)
Eicosanoids
 Key mediators
and regulators of
inflammation and
are generated
from 20 carbon
PUFAs
 Involved in
modulating the
intensity and
duration of
inflammatory
response
(Wymann and Schneiter, 2008)
Utilização de extrato lipídico rico em ARA L.
incisa guppies
Efeito sobre infecção parasitária
(Tetrahymena sp.)
70
Infection (%)
60
50
40
30
20
10
Tetrahymena sp.
0
0.7
12.5
25
50
Arachidonic acid (mg/g)
(Khozin-Goldberg et al., 2006)
21
Efeito sobrevivência de larvas de guppies
da utilização de L. incisa suplementação na
dieta
Cumulative mortality (%)
(via Artemia enrichment)
14
Control
0.1 mg/ml
0.2 mg/ml
0.4 mg/ml
12
10
a
a
8
6
ab
4
Enrichment
(mg ml-1)
ARA
(mg g-1 dw)
Day 34
Lysozyme
(µg g-1 fw)
Day 34
Control
0.1
0.2
0.4
0.8 ± 0.1a
2.0 ± 0.3b
2.9 ± 0.0b
3.2 ± 0.1b
25
38
31
59
2
b
0
0
10
20
30
Time (days)
(Nath et al., 2012)
Parietochloris incisa P127 mutant
Starvation
stage
P. incisa was isolated from a snowy
mountain in Japan
Found to accumulate large amounts of ARA
A mutant, lacking ∆5-desaturase was
produced
P127 - P. Incisa ∆5-desaturase mutant strain,
Accumulates DGLA (20:3n-6)
DGLA, is stored as a triacylglycerol,
within oil globules.
(adopted from Khozin-Goldberg et al., 2011).
Research Approach - Algae
Upscaling and optimizing of DGLA production by P127 stain.
 Optimizing growth conditions:
 Indoor production – Flat PBR (100L; a)
 Outdoor production – Tubular PBR ( 275L; b)
 Optimizing culture parameters
 Initial cell density (growth and starvation phase inoculum)
 Different light intensities
 Mode and quantity of CO2 supply
a
24
b
Desafios do cultivo em larga escala
Contaminacao
Mixing rate
Temperatura
Nutriente
pH
CO2
Luz
25
 Eficiente
 Produção de Composto de Alto
valor
Materiais e métodos
Growth stage culture conditions
Column
Panel
•
•
•
•
•
•
•
P127 cultivated in 2.5 L glass
columns in water baths at 25C
Bubbling of 2% CO2
Light: 180 µmol photons/m2s one
side)
Autoclaved MBG-11
•
PBR panel, room temp.
Bubbling : 2% CO2
Light: 140 µmol photons/m2s each
side
Non autoclaved MBG-11 / MBG-11-N
Sample collection and measurement of growth parameters
- Small sample were analyzed: dry weight, chlorophyll, and for fatty acid
analysis (GC).
Analysis of FA (and DGLA) content by GC
- Direct transmethylated Algal biomass were analyzed by GC method.
Development of P127 upscaled production
Indoor production
1st- Flask 250 mL
2nd- Columns 1 L
3rd- Columns 2.5L
4th- Plastic “V” Bag 15 L
**5th- Flat Photobioreactor(50L/100L)
Growth
stage
Indoor production
**5th- Flat Photobioreactor(50L/100L)
Outdoor production
**5th- Flat Photobioreactor (50L/100L )
6th- Tubular Photobioreactor (275L)
27
Starvation
stage
Cromatografia Gasosa
Materiais e métodos
 Os peixes foram alimentados com
uma
dieta
suplementada
com
diferentes concentrações de P127,
contendo 5%, 10% e 20% e controle
de (w / w)
 Em seguida , os peixes foram
desafiado com Tetrahymena sp. e
novamente amostras foram feitas.
Results - Algae
TFA and DGLA production by P127 under starvation – Flat Panel PBR 50L
%
Time ( Days)
Indoor Flat panel PBR 50 L- initial CHL 30 (optimized conditions)
Objetivos da pesquisa
1. Otimizar a produção de DGLA pela mutante Lobosfera incisa
P127 ( cultivo em larga escala)
2. Avaliar o efeito do uso de suplementação com P127 em dieta de
peixe como imunomodulador em peixe.
Hipóteses:
A produção em grande escala de P127 que
contém níveis elevados de DGLA é viável, mas é
necessária otimização e melhor compreensão do
processo.
Uso de na dieta DGLA pode afetar o status e
doença imune a resistência do peixe.
Cell Breakage and Feed Preparation
Cell
breakage
Dough
• P127 dry algae/ DDW (pH 9)
• Cells were broken by passing
through a Dyano- Mill at 4oC
•Powdered food and algae were
mixed, water added, the dough
was flattened and freeze dried
• The feed was repelled using a
Repelleted steel sieve 500 µm – 1000 µm
Microalgae Biotechnology Laboratory
Prof. Sammy Boussiba - Head
BEA- Banco espanhol de Algas
Lista de Preco de Servicos
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Isolamento de especies
Deposito de patente
Manutencao de cepas
Banco de DNA
 Banco de Cianobacterias
Optimization of TAG and EPA production in one
stage cultivation by light intensity and salinity
management
Variation of cultivation parameters alters the
experimental outcome in Nannochloropsis sp.
Improved biorefinery approaches are required
to take full advantage of the algal biomass
Biorefinery
TAG for
Biofuel
(16:0, 16:1)
High Value
Compounds
(EPA), Protein,
etc.
Pal et al (2011) Appl. Microbiol. Biotechnol.
Cepários Mundiais
Atividades Gerais
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Manutenção de cepas
Venda
Criopreservação
DNA
Educação
Projetos com empresas
Patentes
Isolamentos
Etc
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Culture Collection of Algae at the University of Cologne (CCAC)
Culture Centre for Algae and Protozoa (CCAP)
Sammlung von Algenkulturen Göttingen (SAG)
Chlamydomonas Genetics Center
Canadian Centre for the Culture of Microorganisms (CCCM)
Provasoli-Guillard National Center for Culture of Marine
Phytoplankton (CCMP)
UTEX - The Culture Collection of Algae
Canadian Phycological Culture Centre (CPCC)
CSIRO Collection of Living Microalgae
Microbial Culture Collection-NIES (MCC-NIES)
Cawthron Institute Culture Collection of Micro-Algae (CICCM)
Algobank
Algoteca de Coimbra Culture Collection (ACOI)
Pasteur Culture Collection (PCC)
Roscoff Culture Collection (RCC)
Scandanavian Culture Centre for Algae and Protozoa (SCCAP)
Culture Collection of Autotrophic Organisms ( CCALA )
Culture Collection of Algae of Charles University in Prague
(CAUP)
Culture Collection of Algae of Charles University in Prague
(CAUP)
Plymouth Culture Collection of Marine Microalgae
Sammlung von Algenkulturen Gottingen
(SAG)
BEA- Banco espanhol de Algas
 Projetos
Instalações
Challenges in the Development of
Micro-Algae Based Food Products
Baruch Dach, Co-founder and CTO, AlgaeCore Ltd.
About us
• Founded in 2013
• Develop & Produce micro-algae based staple foods
and specialty chemicals
• Multidisciplinary approach through the whole process
from cultivation to down-stream processing
• “AlgaeMor” Joint Venture (with Mor Group) – Large
scale Micro-Algae R&D & production facility
• Currently engaged in promoting fresh\frozen Spirulina
Product line
www.algae.co.il
Why Bother ?
Shift from Nutraceutical to Functional Foods
Health Food
Going Main Stream
Vegan\ Vegetarian Phenomena
Spirulina
Spirulina (X100)
Spirulina under a SEM
S. Spirulina division (X400)
Spirulina
Spirulina culture
Spirulina biomass
Packaged Spirulina
Questions ? Looking for cooperation ?
[email protected]
AQUANOS
The main cost in wastewater treatment is
the energy needed to oxygenate the
wastewater for the bacteria breaking
down the goo to breathe.
http://aquanos.net/
The Jacob Blaustein Institutes for Desert Research
Obrigada!!
Isabel Campos Portugal
E-mail:
[email protected]
T: +972544539828
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Khozin-Goldberg I., Cohen, Z. Pimenta Leibowitz M., Nechev J., Zilberg D. 2006. Feeding with
arachidonic acid-rich triacylglycerols from the microalga Parietochloris incisa improved recovery of
guppies from infection with Tetrahymena sp. Aquaculture. 255:142-150.
Khozin-Goldberg I., Iskandarov U., and Cohen Z. 2011. LC-PUFA from photosynthetic microalgae:
occurrence, biosynthesis, and prospects in biotechnology. Appl Microbiol Biotechnol. 91:905-15.
Le H. D., Meisel J. A., Meijer V. E., Gura K. M., Puder M. 2009. The essentiality of arachidonic acid
and docosahexaenoic acid. Prostaglandins Leukot Essent Fatty Acids. 81:165-70.
Nath P.T., Khozin-Goldberg I., Cohen Z., Boussiba S., Zilberg D. 2008. Dietary supplementation with
the microalga Parietochloris incisa increases survival and stress resistance in guppy (Poecilia
reticulate) fry. Aquaculture Nutrition. 18:167-180.
Pal D., Khozin-Goldberg I., Cohen Z, Boussiba S. 2011.The effect of light, salinity and nitrogen
availability on lipid production by Nannochloropsis sp. Appl Microbiol Biotechnol. 90:1429-1441
Wang X., Lin H., Gu Y. 2012. Multiple roles of dihomo-g-linolenic acid against proliferation diseases.
Lipid in Heath and Disease. 11:25.
Umeda-Sawada R. et al., 2006. Distribution and Metabolism of dihomo-γ-linolenic acid (DGLA, 20:3
n-6) by oral supplementation in Rats. Biosci. Biotechnol. Biochem. 70(9):2121-2130.