Microbial food colorants

Report
Philippe J. BLANC
Laboratoire Ingénierie des Systèmes Microbiens
et des Procédés
INSTITUT NATIONAL DES SCIENCES APPLIQUEES
135 Avenue de Rangueil
31077 TOULOUSE Cédex 04
FRANCE
Primary identification of food
(tomato = red; lemon = yellow; carrot = orange)
Protection against consumption of spoiled food
Experience, tradition, education, environment
Negative assessment of synthetic food dyes
Strong interest in natural colouring alternatives
An example of traditional food
colorant
Cochineal carmine (red)
OH
O
O 5H12C6
CH3
COOH
HO
OH
OH
O
Others traditional food colorant
Red from paprika, beetroots, berries or tomatoes
Yellow from saffron or marigold
Orange from annatto
Green from leafy vegetables
Acceptability on the market
Regulatory approval
Capital investment size
Carotenoids
Azaphilones
Melanins
Violacein
Flavins
Indigo
Quinones
Monascus pigments: an old
story in Asia but still forbidden
in Europe and the USA
The fungus Monascus
Ang-Kak = Red Rice
Origin = China, Indonesia, Korea,
Japan
Discovered in France by Philippe
van Tieghem in 1884
Lack of toxicity studies
LD50(oral) = 33.3 g/kg (mice)
Not approved in Europe and USA
The chemical structure of the
pigments
C5H11
C7H15
O
O
CH3
O
CH3
O
O
O
O
O
H3C
H3C
O
O
Ankaflavine M = 386
Monascine M = 358
C7H15
C5H11
O
O
CH3
CH3
O
O
O
O
O
O
H3C
H3C
O
O
Monascorubrine M = 382
Rubropunctatine M = 354
C5H11
C7H15
O
O
CH3
O
CH3
O
NH
O
NH
O
H3C
H3C
O
Rubropunctamine M = 353
O
Monascorubramine M = 381
Structure of the free pigments of Monascus
C5H11
O
CH3
O
N
O
COOH
H3C
O
COOH
Glutamylrubropunctatine M = 483
C7H15
O
CH3
O
N
O
COOH
H3C
O
COOH
Glutamylmonascorubrine M = 511
Structure of the complexed pigments of Monascus
The production in various
modes of culture
Red Rice (solid culture)
Liquid culture
The applications
Some advertisements in China (Ningxia
R.D. Natural pigment)
Patents in Japan
- Cosmetic containing pigment derived from
Monascus (Kanebo Lt)
- Monascus culture extract for skin
cleansing and protect effect (Pentel KK)
- Monascus pigment for inhibiting melanin
formation (Hokuto Kasei KK, Ichimaru
Pharmacos Inc.)
Coloration of textiles as silk, wool,
linen (Kasetsart University, Bangkok)
Coloration of biodegradable materials
(ATO/DLO Netherlands)
Others pigments from fungi
Arpink RedTM
an anthraquinonyl derivative
from Penicillium oxalicum
Ascolor Biotech s.r.o.
produced it in
Czech Republic
Fermentation
performances : 2 g/L
Riboflavin from Ashbya gossypii
the vitamin B2 but also a yellow food
colorant
Riboflavin
Ashbya gossypii was originally isolated from cotton as a pathogen !
It produces more than 2 g/L riboflavin
.
b-carotene from various fungi
b-carotene from Blakeslea trispora
DSM was the 1st company to produce b-carotene , today
2 others productions in Russia and Spain.
Others fungi produce b-carotene
Mucor circinelloides
Phycomyces blakesleeanus
Fusarium sporotrichiodes
Astaxanthin from
Xanthophyllomyces dendrorhous
Astaxanthin
Contributes as a flesh pigmenter to the
orange-red coloration of crustaceans and
salmonids
Salmon and trout business is now a huge
business
Fed batch production: 17 g/L biomass
Coloration of crustaceans
Feed non supplemented
with any pigment
Feed supplemented with
astaxanthin
Melanin from Saccharomyces
neoformans var. nigricans
Resarch project presently
Partial conclusion:
Some fermentative food grade
pigments are on the market, many
are at the development stage or
research project
Microbial production of pigments (Industrial production)
Molecule
Colour
Microorganism
Status
Ankaflavin
Yellow
Monascus
IP
Anthraquinone
Red
Penicillium oxalicum
IP
Monascorubramin
Red
Monascus
IP
Rubropunctatin
Purple
Monascus
IP
b-carotene
Yellow-orange
Blakeslea trispora
IP
Riboflavin
Yellow
Ashbya gossypii
IP
Microbial production of pigments (Development stage)
Molecule
Colour
Microorganism
Status
Astaxanthin
Pink-red
Xanthophyllomyces dendrohous
DS
Lycopene
Red
Blakeslea trispora
DS
Rubrolone
Red
Streptomyces echinoruber
DS
Torularhodin
Orange-red
Rhodotorula sp.
DS
Zeaxanthin
Yellow
Flavobacterium sp.
DS
b-carotene
Yellow-orange
Mucor circinelloides
DS
Microbial production of pigments (Research project)
Molecule
Colour
Microorganism
Status
Astaxanthin
Pink-red
Agrobacterium aurantiacum
Paracoccus carotinifaciens
RP
Canthaxanthin
Dark red
Bradyrhizobium sp.
RP
Lycopene
Red
Fusarium sporotrichoides
RP
Melanin
Black
Saccharomyces neoformans var.
nigricans
RP
Naphtoquinone
Deep blood-red
Cordyceps unilateralis
RP
Zeaxanthin
Yellow
Paracoccus zeaxanthinifaciens
RP
b-carotene
Yellow-orange
Fusarium sporotrichiodes
Neurospora crassa
Phycomyces blakesleeanus
RP
Carotenoids from microalgae:
a well established business
Chlorophyceae (green colour)
Rhodophyceae (red colour)
Cyanophyceae (blue green)
Pheophyceae (brown colour)
Dunaliella species for carotenoids
Production: 400 mg b-carotene /m2 of cultivation area
GRAS status
Main companies for carotenoids
Parry’s Agro Ltd (India)
ABC Biotech Ltd. (India)
Betatene Ltd. (Australia)
Western Biotechnology Ltd. (Australia)
Cyanotech Corp., Hawaii (USA)
Inner Mongolia Biological Eng. Co. (China)
Tianjin Lantai Biotechnology (China)
Nature beta Technologies (Israel)
Haematococcus for astaxanthin
Approved by USFDA
Main companies for astaxanthin
Parry’s Pharmaceuticals (India)
Mera Pharmaceuticals, Hawaii (USA)
Cyanotech Co., Hawaii (USA)
BioReal Inc., Hawaii (USA)
Phycoerythrin (fluorescent pink)
from Porphyridium
Production: 200 mg /L of culture
Not yet approved
Phycocyanin (marine blue) from
Spirulina or Porphyridium
Production: 100 mg/L of culture
Not yet approved
Important aspects before selling a new food colorant
Presentation of the microorganism (natural, not GMO)
Lobbying by other colorants producers (nature-identical)
Safety of the fermentation process (HPLC, genotoxicity)
Guidelines for labelling
Conclusions
- Traditional Monascus pigment in Asia
- Success of b-carotene produced by Dunaliella salina
- Success of b-carotene produced by Blakeslea trispora
- Success of Arpink red produced by Penicillium oxalicum
- Exploration of biodiversity to get water-soluble pigments
- Biosynthesis of « niche » pigments not found in plants

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