introduction to microbial pigments used for food

1. Fermentation
and industrial
microbiology
FIM 202

3. Introduction
• A number of different kinds of pigments are produced by
microorganisms. The pigments vary in colour from red, yellow,
orange, purple etc., these pigments play important roles in
photosynthesis, photo protection , pathogenesis etc.
• Microbial pigments have numerous beneficial properties
like anticancer, anti proliferative, immunosuppressive,
antibiotic, biodegradability etc.

4. In food industry
• β –Carotene production
• β –Carotene is a yellowish carotenoid pigments
also known as pro-vitamin A. It acts as
antioxidant and has potential positive properties
against certain diseases.
• Following microbes are
mainly using for β –Carotene production:
• Blakesleatrispora
• Mucorcircinelloides
• Phycomyces blakesleeanus

5. Blakesleatrispora: Some of strains of this mould produce high level of β –
Carotene. B. trispora strains are of two types: (+)mating type and (-)
mating type. Strains obtained by the specific ratio mating of above two
mating types produce β –Carotene14.Today B. trispora fungal β –
Carotene is produced by two industries, the first in Russia and Ukraine,
and the second in Leone, Spain.
Mucorcircinelloides: Wild strains of M.circinelloides, when exposed to the
light impulses of blue light, get activated due to significant change in
structural genes of β–Carotene, hence produce it to high level
Phycomyces blakesleeanus: Phycomycesis mainly used for the production of
various chemicals like β Carotene. They have enhanced carotenogenic
potential when grown on solid substrates or in liquid media

6. • Arpink Red Production
It is the red pigment produced by the strain Penicillium oxalicum
obtained from the soil. It contains chromophore of anthraquinone
type14 .The amounts of red pigment Arpink Red in various food
products was amount recommended by Codex Alimentarius
Commmision.
• Codex Alimentarius Commission (Rotterdam meeting, March
11-15, 2002)
S. No Food product Amount of Arpink Red (in mg/kg)
1. Meat products 100
2. Meat and meat products analogues 200
3. Milk products 150
4. Ice cream 150
5. Confectionery 300

7. • Riboflavin (Vitamin B2) Production
It is the yellow food colourant and its use is permitted in many countries. Because of it specific affinity,
it is used mainly for cereal-based products. Applications of riboflavin somewhere limited due to its bitter
taste and naturally slight odour.
• Different level of Riboflavin production from microbes
• S.No. Producer microorganism Amount produced
1. Weak overproducer 100 mg/L or less Clostridium acetobutylicum
2. Moderate overproducer Upto 600mg/L Candida guilliermundiior
Debaryomyces subglobosus
3. Strong overproducer Over 1g/L Eremothecium ashbyiiand, Ashbyagossypi
Fermentation with Ashbyagossypi is preferred because of higher yield and greater genetic stability;
riboflavin levels of over 15g/L have been reported

8. • Monascus pigments production
Monascus spp. belongs to the group of Ascomycetes and particularly to
the family of Monascaceae. The genus Monascus can be divided
into four species: M. pilosus, M.purpureus, M ruberand M. froridanus,
which account for the majority of strains isolated from traditional
oriental food.
• The common names of this fungal product are Red Yeast Rice (RYR), red
rice, angkak, red leaven, benikoji (Japanese), hung-chu, hongqu, zhitai
(Chinese), rotschimmelreis (Europe), red mould (USA) and MFR
(Monascus fermented rice).
• Monascus spp. produce many pigments of industrial importance and
these
pigments are mainly of three types i.e. red colorants, orange colorants
and yellowish colorants.
• Monascus fungi, organisms produce angkak can convert starchysubstrates
into several metabolites such as alcohols, antibiotic agents,
antihypertensives enzymes, fatty acids, flavor compounds, flocculants,
ketones, organic acids, pigments and
vitamins. Thus, the implementation of Monascus pigment as a coloring
agent in food provided an
additional advantage of specific flavor in the
products.

9. • Lycopene Production
It is a red open-chain unsaturated carotenoid, acyclic
isomer of beta-carotene, and longer than any other
carotenoid .
• Lycopene, also known as psi-carotene, is very
sensitive to heat and oxidation and is insoluble in
water, Because of the abundance of double bonds in
its structure.
• Genetically modified fungus
Fusarium sporotrichioides was used by Jones etal to
manufacture the colourant and antioxidant lycopene.
They used the cheap corn fiber material as the
substrate.
• Cultures in lab flasks produced 0.5 mg (lycopene)/g
of dry mass within 6 days and such a production will
be increased within the next years.

10. Microorganism(s) Pigments/Molecule Colour/appearance
Bacteria
Agrobacterium aurantiacum Astaxanthin Pink-red
Paracoccus carotinifaciens Astaxanthin Pink-red
Bradyrhizobium sp. Canthaxanthin Dark- red
Flavobacterium sp.,
Paracoccus zeaxanthinifaciens Zeaxanthin yellow
Achromobacter Creamy
Bacillus Brown
Brevibacterium sp. Orange yellow
Corynebacterium michigannise Greyish to creamish
Corynebacterium insidiosum Indigoidine Blue
Rugamonas rubra , Streptoverticillium rubrireticuli,Vibrio
gaogenes, Alteromonas rubra
Prodigiosin Red
Rhodococcus maris Bluish- red
Xanthophyllomyces dendrorhous Astaxanthin Pink -red
Haloferax alexandrinus Canthaxanthin Dark Red
Staphylococcus aureus Staphyloxanthin Zeaxanthin Golden Yellow
Chromobacterium violaceum Violacein Purple
Serratia marcescens, Serratia rubidaea, Prodigiosin Red
Pseudomonas aeruginosa Pyocyanin Blue-green
Xanthomonas oryzae Xanthomonadin Yellow
Janthinobacterium lividum Violacein Purple

11. Algae
Dunaliella salina β-carotene Red
Chlorococcum Lutein
Hematococcus Canthaxanthin
Fungi
Aspergillus sp. Orange-red
Aspergillus galucus Dark -red
Blakeslea trispora β -carotene Cream
Helminthosporium catenarium Red
Helminthosporium avenae Bronze
Penicilllum cyclopium Orange
Penicilllum nalgeovensis Yellow
Fusarium sporotrichioides Lycopene Red
Haematococcus Pluvialis Astaxanthin Red
Monascus sp. Monascorubramin Rubropunctatin Red Orange
Monascus purpureus Monascin Ankaflavin Red-yellow
Monascus roseus Canthaxanthin Orange-Pink
Monascus sp. Ankaflavin Yellow
Penicillium oxalicum Anthraquinone Red
Blakeslea trispora Lycopene Red
Cordyceps unilateralis Naphtoquinone Deep blood-red
Ashbya gossypi Riboflavin Yellow
Mucor circinelloides, Neurospora crassa and Phycomyces
blakesleeanus

12. • Benefits and Applications of Microbial pigments
• Pigments produced by microorganisms are of traditional use in oriental
countries and have been a subject of intense research in the present
decades because of its potential for applications. Use of microorganisms
and biotechnology would offer solutions to the problems of various
industries especially food colorant industry.
• Fermentative production of colorants has a number of benefits that
include: cheaper production, readily available raw materials, high yields
and no seasonal variations . In contrast to higher plants, single cell algae
and fungi are more appropriate for biotechnological production because
they can be grown using existing culture techniques. Many fungi produce
pigments which have application in both textile and food industries .
• Fungal pigments are routinely utilized as colorants for both foodstuffs
and materials. Some of the more importantly utilized fungal pigments
come from the water soluble orange/red pigments produced by
Monascus sp., frequently used in rice wines in eastern countries .
Pigments produced from Monascus purpureus Piedallu are used in wool
dying , and an anthraquinone pigment obtained from Penicillium
oxalicum Currie & Thom is currently being developed for use as a ‘natural’
food additive that may have some anticancer effects.

13. • Mycelial extracts of some promising mushrooms are Chroogomplus vinicolor
(which gives red tints), Bankera violascens (which gives greens) and Collybia iocephala
(which gives blues), they have a remarkable potential for dyeing wool and silk fabrics.
Carotenoids such as β-carotene and lycopene have been known to be produced by fungal
cell factories.
• For example, β-carotene by Blakeslea trispora, Mucor circinelloides, and
Phycomyces blakesleeanus and lycopene from Fusarium sporotrichioides, Blakeslea
trispora are already in use as for food colorants . Vitamin, riboflavin (vitamin B2) is a
yellow.
• food colorant that is fermentatively produced by the fungi Eremothecium ashbyii and
Ashbyagossypi and has huge commercial potential . Microalgae such as Haematococcus
(astaxanthin), Spirulina (phycocyanin), and Dunaliella (ß-carotene and other carotenoids)
are already explored as sources of natural food colorants with some limitations of low
productivity and contamination in the open culture system, where they are grown [76].
Newly, a blue-green polyphenolic antioxidant pigment termed marennine from the
Diatom Haslea ostrearia has been reported . This may aid as a promising food coloring
additive. Among yeasts, the marketable success has been with the case of astaxanthin
production by Xanthophyllomyces dendrorhous (formerly Phaffia rhodozyma) and is
widely in use as food additive. Additional example of production of carotenoids is by the
yeast Rhodotorula spp., R. glutinis, R. gracilis, R. rubra, and R. graminis. The chief
carotenoids produced by these yeasts are torulene and torularhodin, with minute
quantity of β-carotene .

14. • The pigments obtained from bacteria offer the following benefits and advantages ;
• they are progressively attractive to science because of broad ranging activities,
easy propagation and wide-ranging strain selection, high versatile and productive
compared to other sources, fermentation is integrally faster and more productive
production when compared to any other chemical process, simple and fast
culturing methods allowing continuous bioreactor operation, quite easy to
manipulate genes, structural complexity suits for industrial requirements, Cheap
substrates used for bulk production and their pigments extracted using simple
liquid-liquid extraction technique minimizing operation cost.
•

15. • Some important functions of bacterial pigments of commercial potential are
• 1) Riboflavin synthesized by Bacillus subtilis and used in foods, vitamin enriched milk products and
energy drinks, 2) β-carotene usually produced by Flavobacterium, Agrobacterium aurantiacum and
widely used as food supplement for humans and as food additives for animals and fish,
• 3) Prodigiosin by Serratia marcescens, Pseudomonas, Pseudoalteromonas, Alteromonas
denitrificans, Hahella, Vibrio species used as antibacterial, antifungal, antimalarial, antibiotic,
immunosuppressive, anticancer etc.,
• 4) Phycocyanin by Cyanobacteria used as dietary supplement rich in proteins, and
• 5) Violacein by Chromobacterium violaceum,Alteromonas luteoviolacea was used as Antiviral,
antibacterial, antiulcerogenic, antileishmanial, and anticancer properties, potent cytotoxic effects
against U937 and HL60 Leukemia cell lines

16. Novel practices of microbial pigment production
• Strain improvement
• Fermentation
• Low-cost substrates