A study on the conversion of carotene into vitamin A by fluorescence microphotometry

For the purpose to confirm whether carotene is converted into vitamin A mainly in the intestine, fluorescence microscope observations as well as the fluorescence microphotometry for the estimation of fading state of the fluorescence were carried out on the fresh sections of the intestine, lung and liver of rats after oral administration of 11-carotene dissolved in oil, and suspended in water, and vitamin A in sesame oil as control. Yellowsih-green fluorescence of carotene is similar to that of vitamin A in color but the fluorescence of vitamin A fades away very rapidly within one minute while that of carotene does not fade or fade more slowly than that of vitamin A. Observations have revealed that, contrary to expectation, the administered carotene is not so readily converted into vitamin A in the intestinal mucosa, but after passing through the intestine without conversion to vitamin A, it is transported to the mesenteric lymph vessels, portal vein, and reaches the liver. In the liver, carotene appears as fatty droplets or micronized particles in the parenchymal cell. The conversion of carotene into vitamin A could not be observed in the intestine, liver and lung in the observations made one hour after the oral adminstration of carotene. Hower, it seems that carotene dissolved in minute fat droplets may be converted into vitamin A at water phase in tissues, after dissolution of carotene in fat and micronization of the fat droplets.</p

ENHANCED METHOD FOR AMPLIFYING ANTIMICROBIAL ACTIVITY OF LACTOPEROXIDASE SYSTEM IN MILK AND DERIVED PRODUCTS BY CARROT EXTRACT AND BETA CAROTENE

OSCN–  has  been  known  and  well  documented  as  natural  antimicrobial  agent    generated  from the  lactoperoxidase  system (LPOS)  but  the  antimicrobial  activity  exerted  from  this  system  is  too  low  for  certain  food  such  as  milk  and  derived products (up to 1.2 log CFU/ml) resulting in the restriction on industrial  development  of  LPOS.  Our  previous  study  concluded  that  involvement  of  carrot  extract and beta carotene in LPOS significantly boosted the antimicrobial activity  (up  to  6  log  CFU/ml)  against  S.  enteritidis.  This  finding  should  be  continued  to  the  application  on  food.  Since  we  found  that  LPOS  generates  low  antimicrobial  activity  on  milk  and  derived  product,  our  research  will  be  conducted  on  the  application  of  LPOS  plus  carrot  extract  and  beta  carotene  on  milk  and  derived  product.  Because  the  high  antimicrobial  activity  is  needed  for  industrial  purposes,  this  research  may  open  the  way  for  industrial  development  of  natural  antimicrobial agent from LPOS.  This research will be conducted in three steps of experiment: (1) ensuring  the  incredible  antimicrobial  activity  of  LPOS  plus  carrot  extract  and  beta  carotene  against  three  poisonous  tropical bacteria:  C. jejuni,  S. enteritidis,  and E.  coli,  (2)  utilization  of  LPOS  plus  carrot  extract  and  beta  carotene  in  milk:  full  cream and skimmed milk, and (3) utilization of LPOS plus carrot extract and beta  carotene  in  milk  derived  product:  yogurt  and  tropical  fruity  milk.  The  purification  of  LPO  from  bovine  milk,  the  complimentary  data  analysis  for  generation  of  LPOS  antimicrobial  activity  by  the  addition  of  carrot  extract  and  beta  carotene  will  be  conducted  in  Japan  and  for  the  application  in  tropical  poisonous  bacteria,  milk,  and  milk  derived  product  will  be  conducted  in  Indonesia.  The  outcomes  of  this  research  from  three  step  of  research  are  three  international  publications  at  international‐scopus‐indexed‐journals  :  International Journal of  Dairy Science and  one  of  patent:  “enhanced  method  for  improvement of LPOS antimicrobial activity by involvement of carrot extract and  beta carotene in milk and derived product”.     Keywords:  Lactoperoxidase,  antimicrobial  activity,  carrot  extract,  beta  carotene,  milk, derived products

The carotenoid and provitamin A content of the watermelon

10 years ago lycopene, C40H56, and carotene, C40H56, were isolated from the pulp of the European watermelon (Citrullus vulgaris, Schrad. = Cucumis citrullus, L.) by Zechmeister and Tuzson (l), and it was shown that the chief pigment, lycopene, is responsible for the red color. As the chromatographic method was not available at that time, no precise information as to the composition of the pigment was obtained. In some new experiments described below we have carried out a quantitative analysis of the components and have estimated the provitamin A content of the California watermelon. 1 kilo of the pulp examined contained 1.0 mg. of a complicated xanthophyll mixture, 6.1 mg. of lycopene, 0.06 mg. of γ-carotene, 0.16 mg. of unknown carotenoids (located in the column between γ- and β-carotene), 0.46 mg. of β-carotene, 0.01 mg. of α-carotene. The figures include the fractions of lycopene, and γ- and β-carotene which underwent isomerization during the experimental procedure (2). The calorimetric value of the total extract of 1 kilo of pulp corresponded to 7 to 8 mg. of “lycopene”; some samples were, however, considerably richer in pigment. Our material, picked in California in September, contained, according to the above figures, 0.5 mg. of provitamin A in 1 kilo of pulp, or about one-fifth to one-sixth of the daily β-carotene requirement of an adult person. It is interesting to note that a considerable number of yellow and pink unidentified oxygen-containing carotenoids were found in minute quantities; i.e., to the extent of about 0.01 mg. per kilo of pulp. Even with the use of chromatography 1000 or more kilos of melon would be needed for a satisfactory study of these pigments. One of them is spectroscopically identical with torulene, detected by Lederer in red yeast (3)

Genes involved in carotene synthesis and mating in Blakeslea trispora

Mating of Blakeslea trispora and other molds of the order Mucorales requires the interaction of mycelia of opposite sex, (+) and (-), leading to the development of specialized structures and to an enhanced accumulation of beta-carotene. Industry obtains beta-carotene by co-cultivating appropriate strains of Blakeslea (mated cultures). Gene transcription in single and mated cultures was assayed by cDNA-AFLP, a technique to observe the differential expression of subsets of mRNA fragments. Overexpression in mated cultures is about ten times more frequent than underexpression. We obtained and sequenced fragments of 97 candidate genes that appeared to be overexpressed during mating and confirmed four of them by reverse transcription and real-time PCR. Comparisons with gene sequences from other organisms suggest functions in carotene biosynthesis (4 genes), energy metabolism (8), cell wall synthesis (1), transfer of acetyl groups (1), and regulatory processes (10). Sodium acetate inhibited sexual overexpression in about two-thirds of the candidate genes and acted as a signal with broad effects on the metabolism and the morphology of mated cultures. Our work offers new materials for the study of carotene biosynthesis and its regulation and for the improvement of carotene production with Mucorales

Elaboration of New Method of Deep Processing of Caro-tene-containing Raw Materials Into Nanoadditives with the Use of Cryogenic Freezing and Fine-dispersed Grinding

The aim of the work is elaboration of principally new cryogenic method of deep processing of carotene-containing vegetable raw material (CCVRM) such as carrot, pumpkin, sweet Bulgarian pepper, tomato, sea buckthorn, apricot using cryogenic freezing and fine-dispersed grinding.The new method of deep processing leads to more full extracting and removal of β-carotene from the state, bound with biopolymers into free and hydrophilic form. The mechanism of these processes was presented. It was established, that mass share of β-carotene in cryopuree, received by the new method, exceeds its content in initial (fresh) CCVRM in 3,0…3,5 times.The regularities of growth and transformation of carotenoids separately at freezing with different high speeds and low-temperature grinding of CCYRM were established. It was demonstrated, that at cryogenic freezing take place quantitative increase of mass share of carotenoids in 2,0…2,5 times comparing with initial (fresh) raw material depending on freezing speed and type of CCVRM. That is it was demonstrated, that frozen carotene-containing vegetable raw material contains 2,0…2,5 times more β-carotene than fresh one. Mechanism of these processes was presented.It was demonstrated, that cryopuree of CCVRM, received using new methods of deep processing, exceeds the initial (fresh) raw material by the content not only β-carotene but also other low-molecular bioactive substances: L-ascorbic acid – in 2,0…2,2 times, phenol compounds – in 1,7…1,8 times, tanning substances – in 1,5…1,7 times). That is they have principally new chemical composition, because practically all bioactive substances (BAS) in cryopuree are in nanosize form and easily assimilated by human organism

Improving the in vitro bioaccessibility of β-carotene using pectin added nanoemulsions

The intestinal absorption of lipophilic compounds such as β-carotene has been reported to increase when they are incorporated in emulsion-based delivery systems. Moreover, the reduction of emulsions particle size and the addition of biopolymers in the systems seems to play an important role in the emulsion properties but also in their behavior under gastrointestinal conditions and the absorption of the encapsulated compound in the intestine. Hence, the present study aimed to evaluate the effect of pectin addition (0%, 1%, and 2%) on the physicochemical stability of oil-in-water nanoemulsions containing β-carotene during 35 days at 4 °C, the oil digestibility and the compound bioaccessibility. The results showed that nanoemulsions presented greater stability and lower β-carotene degradation over time in comparison with coarse emulsion, which was further reduced with the addition of pectin. Moreover, nanoemulsions presented a faster digestibility irrespective of the pectin concentration used and a higher β-carotene bioaccessibility as the pectin concentration increased, being the maximum of ≈36% in nanoemulsion with 2% of pectin. These results highlight the potential of adding pectin to β-carotene nanoemulsions to enhance their functionality by efficiently preventing the compound degradation and increasing the in vitro bioaccessibility.This work was funded by the project AGL2015-65975-R (FEDER, MINECO, UE) and project RTI2018-094268-B-C21 (MCIU, AEI; FEDER, UE)

Application of near infrared reflectance spectroscopy in screening of fresh cassava (Manihot esculenta crantz) storage roots for provitamin A carotenoids

A developed Near Infrared Reflectance Spectroscopy (NIRS) calibration equation was used for determining provitamin A carotenoids contents of different trials of fresh yellow root cassava genotypes using a total of 50 cassava genotypes scanned twice by NIRS from 400 nm to 2498 nm. The NIRS calibration equations were used to predict the β-cryptoxanthin, 13-cis β-carotene, trans β-carotene, 9-cis β -carotene, total β-carotene and total carotenoid concentrations of the samples. The predicted values for total carotenoids (TC-pred) ranged from 3.93 μg g–1 to 10.51 μg g–1 with mean of 7.07 ± 2.55 μg g–1 for International Collaborative Trials (ICT), 7.97–11.03 μg g–1 fresh weight with mean of 9.40 ± 0.76 μg g–1 for yellow root trial 8 (Multi-location Uniform Yield Trial) and 6.38–10.44 μg g–1 with mean of 8.74 ± 1.07 μg g–1 for yellow root trial 9 (Multilocation Advanced Yield Trial). Total carotenoids results using reference spectrophotometric method (TC-spec) ranged from 2.57 μg g–1 to 9.97 μg g–1 with mean of 5.66 ± 2.99 μg g–1 for ICT, 6.55–8.74 μg g–1 with mean of 7.74 ± 0.64 μg g–1 for yellow root trial 8 and 4.22–11.00 μg g–1 with mean of 7.57 ± 1.54 μg g–1 for yellow root trail 9. There is significant (P ≤ 0.001) positive correlation (r = 0.55) between TC-pred by NIRS and TC-spec. Also, significant (P ≤ 0.001) positive correlation (r = 0.52) exist between trans β-carotene predicted by NIRS and high-performance liquid chromatography reference. The developed NIRS calibration equations could be used to predict total carotenoids and trans β-carotene content of yellow root cassava and serve as rapid and cost-effective screening method for large cassava sample sets