TABLE 3.

Characteristics of carotenoids extracted from the wild type and the mutants and the corresponding genetically modified Ha. japonica strains

Strain	Peak no.	Retention time (min)	λmax (nm) in HPLC eluent	Conjugated double bonds (n)	Molecular mass (m/z)		Carotenoida
					Measured	Calculated
Ha. japonica WT	1	9.6	469, 492, 525	13	740	740	BR
	2	11.8	469, 491, 522	13	722	722	MABR
	3	13.8	467, 490, 521	13	704	704	BABR
	4	16.0	453, 479, 509	12	620	620	IDR
	5	19.4	442, 468, 499	11	536	536	Lycopene
Ha. japonica Δc0506	5	19.4	443, 469, 500	11	536	536	Lycopene
Ha. japonica Δc0507	4′	14.8	441, 466, 498	11	708	708	TH-BABR
	5′	16.7	440, 465, 495	11	622	622	DH-IDR
Ha. japonica Δc0505	3	13.7	468, 491, 523	13	704	704	BABR
Ha. japonica Δc0506(pJc0506)	1	9.6	469, 491, 523	13	ND	740	BR
	2	11.8	468, 491, 523	13	ND	722	MABR
	3	13.8	466, 490, 522	13	ND	704	BABR
Ha. japonica Δc0507(pJc0507)	1	9.6	468, 492, 524	13	ND	740	BR
	2	11.8	469, 491, 522	13	ND	722	MABR
	3	13.8	466, 490, 521	13	ND	704	BABR
	4	16.0	454, 480, 510	12	ND	620	IDR
	5	19.4	443, 468, 499	11	ND	536	Lycopene
Ha. japonica Δc0505(pJc0505)	1	9.6	468, 492, 525	13	ND	740	BR
	2	11.8	467, 492, 522	13	ND	722	MABR
	3	13.8	466, 490, 520	13	ND	704	BABR
	4	16.0	454, 479, 509	12	ND	620	IDR

^aBR, bacterioruberin; MABR, monoanhydrobacterioruberin; BABR, bisanhydrobacterioruberin; IDR, isopentenyldehydrorhodopin; TH-BABR, tetrahydrobisanhydrobacterioruberin; DH-IDR, dihydroisopentenyldehydrorhodopin. Carotenoids were analyzed using an HPLC system equipped with a μBondapak C₁₈ column (Waters) (3.9 by 300 mm) and were eluted with methanol-water (9:1 [vol/vol]) for the first 10 min and then with 100% methanol (1.5 ml/min). Absorption spectra of the carotenoids were recorded with a photodiode array detector attached to the HPLC apparatus (Fig. 3). The relative molecular masses of the purified carotenoids were measured using an MStation JMS-700 mass spectrometry system (Jeol) in the FAB mode with m-nitrobenzyl alcohol as a matrix. ND, not determined.