Mutant chlorella strain capable of producing zeaxanthine and beta-carotene and culturing method thereof

A technology of carotene and zeaxanthin, applied in the field of microbial engineering

Active Publication Date: 2017-03-22
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] So far, there is no report in the prior art of a mutant strain ZEA1 that

Method used

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  • Mutant chlorella strain capable of producing zeaxanthine and beta-carotene and culturing method thereof
  • Mutant chlorella strain capable of producing zeaxanthine and beta-carotene and culturing method thereof
  • Mutant chlorella strain capable of producing zeaxanthine and beta-carotene and culturing method thereof

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Embodiment 1

[0038] 1) Inoculate the starting algal strain Chlorella (Chlorella zofingiensis, ATCC 30412) (American TypeCulture Collection, ATCC, Rockville, MD, USA) from Kuhl agar slant medium (containing 3g / L glucose and 1.5% agar) to Cultured for 4 days in 10ml of Kuhl's liquid medium. The culture conditions were as follows: temperature 25 degrees Celsius, shaker speed 150 rpm, light intensity 30umol m -2 the s -1 . The composition of Kuhl medium is (L): 1.01g KNO 3 ; 0.62gNaH2PO4 H2O; 0.089g Na 2 HPO 4 2H 2 O; 0.247g MgSO 4 ·7H 2 O; 14.7 mg CaCl 2 2H 2 O; 6.95 mg FeSO 4 ·7H 2 O; 0.061 mg H 3 BO 3 ; 0.169 mg MnSO 4 ·H 2 O; 0.287mg ZnSO 4 ·7H 2 O; 0.0025 mg CuSO 4 ·5H 2 O; and 0.01235mg (NH 4 ) 6 MO 7 o 24 4H 2 O; pH 6.8.

[0039] 2) Inoculate the algae cells cultured in the above bottle for 4 days into fresh 50 ml Kuhl liquid medium at a ratio of 1:10 and culture them for 4 days.

[0040] 3) Collect the algae cells after 4 days of culture in step 2) by centrifu...

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Abstract

The invention provides a mutant chlorella strain capable of realizing high yield of zeaxanthine and other carotenoids essential to the human body, and a culturing method thereof. The culturing method comprises the following steps: subjecting Chlorella zofingiensis ATCC 30412 to nitrosoguanidine mutagenesis; then coating a Kuhl solid medium containing DPA with treated chlorella cells for culture; selecting a single chlorella colony from the Kuhl solid medium and culturing the single chlorella colony on a Kuhl liquid medium; adding glucose in the middle and later periods of logarithm for induction of carotenoid synthesis; and detecting the variety changes and content of carotenoids in the obtained chlorella body so as to obtain the strain which can realize high yield of zeaxanthine, xanthophyll and beta-carotene and is named as CZ-ZEA1. Under the induction condition that the concentration of glucose is 30 g/L, the biomass of the CZ-ZEA1 is 12.95 g/L, and the contents of zeaxanthine, xanthophyll and beta-carotene reach 2.176 mg/g, 1.10 mg/g and 1.211 mg/g, respecitively. The mutant chlorella strain is an edible alga species and contains high-content optimally-proportioned carotenoids essential to the human body, especially zeaxanthine rare in nature; so the mutant chlorella strain is an ideal new-resource functional foodstuff for preventing deterioration and lesion of the eyes of middle aged and old people and has good development and application prospects.

Description

technical field [0001] The invention belongs to the field of microbial engineering, and in particular relates to a mutant chlorella strain with high yield of zeaxanthin and beta-carotene produced by nitrosoguanidine mutagenesis and a cultivation method thereof. [0002] technical background [0003] Zeaxanthin is a yellow hydroxylated carotenoid, its scientific name is 3,3'-dihydroxy-β-carotene, and like β-carotene, zeaxanthin has strong coloring and antioxidant functions . Zeaxanthin and its isomer lutein are the main pigment components of the macula of the eye, and their content ratio in the macula is 2:1. Studies in recent years have shown that zeaxanthin helps prevent cataracts, age-related macular degeneration, cancer, and low-density lipoprotein oxidation. In addition, zeaxanthin is also widely used in poultry and fish farming as a feed additive. At present, the main source of zeaxanthin on the market is biological extraction, and natural sources of zeaxanthin are mai...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N15/01C12R1/89
CPCC12N15/01C12N1/125C12R2001/89
Inventor 黄俊潮黄为平林燕
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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